Sexual reproduction and short-term fitness advantage in the rotifer Brachionus calyciflorus: implications for the coexistence of sympatric clones

Large numbers of rotifer eggs from sediment resting egg banks may hatch simultaneously under appropriate conditions;therefore,natural populations are likely to be multiclonal in a growing season.A recent field investigation showed that subordinate and ephemeral clones were able to establish populations in an environment with several strongly dominant clone populations.However,it was not clear how the subordinate populations maintained their growth under these conditions where the crowding signal would induce high levels of sexual reproduction in the dominant clone populations.In the present study,we conducted a continuous passage to 60 generations for new populations at three different temperatures(15°C,25°C,and 35°C).These populations emerged from resting eggs produced by a clone population.At the first,30th,and 60th generations,the populations were sampled and fitness was assayed using the life-table method.We found a significant short-term fitness advantage for populations newly emerged from resting eggs produced by a clone population of the rotifer Brachionus calyciflorus,followed by a significant decrease in relative fitness with long-term continuous passage.Our results suggested that the early shortterm fitness advantage was important for later-hatched and subordinate clone populations in an environment with multiple sympatric clones.The early short-term fitness might also promote the coexistence of sympatric clones because it could help to offset the cost of early sexual reproduction.     

Effects of sexual reproduction of the inferior competitor Brachionus calyciflorus on its fitness against Brachionus angularis

Sexual reproduction adversely affects the population growth of cyclic parthenogenetic animals. The density-dependent sexual reproduction of a superior competitor could mediate the coexistence. However,the cost of sex may make the inferior competitor more vulnerable. To investigate the effect of sexual reproduction on the inferior competitor,we experimentally paired the competition of one Brachionus angularis clone against three B rachionus calyciflorus clones. One of the B. calyciflorus clones showed a low propensity for sexual reproduction,while the other two showed high propensities. The results show that all B. calyciflorus clones were excluded in the competition for resources at low food level. The increased food level promoted the competition persistence,but the clones did not show a clear pattern. Both the cumulative population density and resting egg production increased with the food level. The cumulative population density decreased with the mixis investment,while the resting egg production increased with the mixis investment. A trade-off between the population growth and sexual reproduction was observed in this research. The results indicate that although higher mixis investment resulted in a lower population density,it would not determinately accelerate the exclusion process of the inferior competitor. On the contrary,higher mixis investment promoted resting egg production before being excluded and thus promised a longterm benefit. In conclusion,our results suggest that mixis investment,to some extent,favored the excluded inferior competitor under fierce competition or some other adverse conditions.     

Reduction of hydrocarbon contamination on viability of Acartia pacifica benthic resting eggs

The potential effect of hydrocarbon contamination on the hatching success of benthic resting eggs of Acartia pacifica in Xiamen Bay was investigated experimentally. The number of nauplii emerging from the sediment samples decreased with increasing Fuel Oil #0 concentration. The estimated rate of mortality increased markedly with the increase of Fuel Oil #0 concentration. Successive fuel Oil #0 concentrations from 50 mg/kg to 5000 mg/kg reduced the number of hatched nauplii by 3.8% -100%. The mortality of A. pacifica resting eggs due to Fuel Oil #0 contamination did not significantly increase as time progressed at each concentration level. The LC50 values of resting eggs, changing from 237.12 to 279.59 mg/kg, remained at an almost stable level in two months. The number of A. pacifica nauplii that hatched from the sediment at 10℃ was higher than those from the sediment at 30℃, which indicates that the toxicity of Fuel Oil #0 on A. pacifica resting eggs increases with increasing temperature.     

Comparison of two sampling methods when studying periphyton colonization in Lam Tsuen River, Hong Kong, China

We used standard periphyton samplers to examine the colonization pattern of periphytic algae on artificial substrates (glass slides) in Lam Tsuen River, Hong Kong, in dry (winter) and wet (summer) seasons. In each season, six replicated slides were retrieved randomly and replaced by new slides at weekly intervals over a period of 6 weeks. We thus obtained two batches of slides, both with a series of different exposure times (1 to 6 weeks): one batch was set up at the same time (start of the sampling) and the other was harvested at the same time (end of the sampling). Changes in taxonomic composition, species diversity (Shannon-Wiener diversity index), standing crop (in terms of cell density and cell biovolume), and abundance of the abundant algal species were monitored and compared between the two batches of slides. The succession patterns of the periphytic algae were similar between the two batches in each season, while more remarkable differences were observed between the two seasons, suggesting that either batch would be suitable for a colonization study of periphytic algae. The cell density was dominated by diatoms in both seasons, while the cell biovolume was dominated by diatoms in winter and by green algae in summer. The cell density and biovolume attributed to blue green algae was relatively small. Most of the diatom species exhibited similar colonization patterns throughout the experiment, while green algae showed different succession patterns in different seasons or sampling methods, indicating that diatoms are better bio-indicators than green algae for a periphyton colonization study. In general, the diversity indices and the standing crops reached their maximums at around week 4, and they were higher in summer than in winter.     

Temperature Effect on the Life History of Three Types of Brachionus calyciflorus Females

The effect of temperature on the life history characteristics of amictic females ( AF), unfertilized mictic females (UMF) and fertilized mictic females (FMF) in Brachionus calyciflorus was studied with replicated individual cultures at 20℃ ,25℃ and 30℃ , and with algae Scenedesmus obliquus for their food. There were highly significant effects of both temperature and female type, independently and interactively, on the duration of juvenile and post-reproduction periods, and the number of eggs produced by the rotifer per life cvcle. Among all the temperature-female type combinations, all the juvenile periods of FMF, and the post-reproduction periods of UMF and FMF at 20℃, were the longest, and the number of eggs produced by an UMF at 30℃ was the highest. There were highly significant effects of both temperature and female type on the duration of the reproduction period, but no clear correlation was observed between temperature and female type. The reproduction period of AF was longer than that of UMF and FMF.Only temperature influenced significantly the mean life-span of the three types of females. The duration of juvenile,reproduction and post-reproduction periods as well as the life-span of the three types of females were all reduced very significantly with rise of temperature, but the rates of reduction varied with female type. Among the three types of females, the number of eggs produced per life cycle by an UMF was the highest, and that of a FMF was the lowest. A significant relationship between the number of eggs produced per life cycle and temperature was observed only in the UMF.     

Construction of a Metagenomic DNA Library of Sponge Symbionts and Screening of Antibacterial Metabolites

To study the bioactive metabolites produced by sponge-derived uncultured symbionts, a metagenomic DNA library of the symbionts of sponge Gelliodes gracilis was constructed. The average size of DNA inserts in the library was 20 kb. This library was screened for antibiotic activity using paper disc assaying. Two clones displayed the antibacterial activity against Micrococcus tetragenus. The metabolites of these two clones were analyzed through HPLC. The result showed that their metabolites were quite different from those of the host E. coli DH5α and the host containing vector pHZ132. This study may present a new approach to exploring bioactive metabolites of sponge symbionts.     

High genetic diversity in gametophyte clones of Undaria pinnatifida from Vladivostok, Dalian and Qingdao revealed using microsatellite analysis

Breeding practice for Undaria pinnatifida (Harvey) Suringar requires the screening of a large number of offspring from gametophyte crossings to obtain an elite variety for large-scale cultivation. To better understand the genetic relationships of different gametophyte cultures isolated from different sources, 20 microsatellite loci were screened and 53 gametophyte clone cultures analyzed for U. pinnatifida isolated from wild sporophytes in Vladivostok, Russia and from cultivated sporophytes from Dalian and Qingdao, China. One locus was abandoned because of poor amplification. At the sex-linked locus of Up-AC-2A8, 3 alleles were detected in 25 female gametophyte clones, with sizes ranging from 307 to 316 bp. At other loci, 3 to 7 alleles were detected with an average of 4.5 alleles per locus. The average number of alleles at each locus was 1.3 and 3.7 for Russian and Chinese gametophyte clones, respectively. The average gene diversity for Russian, Chinese, and for the combined total of gametophyte clones was 0.1, 0.4, and 0.5, respectively. Russian gametophyte clones had unique alleles at 7 out of the 19 loci. In cluster analysis, Russian and Chinese gametophyte clones were separated into two different groups according to genetic distance. Overall, high genetic diversity was detected in gametophyte clones isolated from the two countries. These gametophyte cultures were believed to be appropriate parental materials for conducting breeding programs in the future.     

Dark touristic perception: Motivation,experience and benefits interpreted from the visit to seismic memorial sites in Sichuan province

This study investigates the motivations of Chinese domestic tourists visiting seismic memorial sites after the great Wenchuan earthquake of 2008, and examines their interpretation of experiences and benefits of the visits. Recent research on dark tourism has raised the possibility that people are attracted to death and memorial sites in ways different from other tourist attractions. The study used an empirical research design employing questionnaires to collect data from 255 Chinese domestic visitors at seismic memorial sites in west Sichuan. Analysis revealed that the obligation of commemoration mixed with curiosity represents a different set of travel motivation in dark tourism settings than in other kinds of tourism. In addition, the empirical evidence also suggests an interrelated pattern between motivations, experiences and benefits. This study implies that Chinese domestic tourists were attracted in ways different from other tourist attractions both because they tended to fulfill their obligation of commemoration and were interested in destruction; and visitor experiences played an important mediating role between travel motivations and benefits gained.     

STUDY ON MIXIS POTENTIAL OF ROTIFER RESTING EGGS ( BRACHIONUS PLICATILIS ) WITH DIFFERENT COLLECTION TIMES AND DIFFERENT PRESERVATION PERIODS

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Triploidy induction with heat shocks toPenaeus chinensis and their effects on gonad development

Heat shocks effectively produced triploids inPenaeus chinensis. Fertilized eggs heat shocked (28–32°C) for 8 to 16 minutes, starting from 8 to 20 minutes after spawning, resulted in triploidy induction rates of 39%–75%. Several triploid populations were cultured to 10 cm. In a triploid population, two kinds of ovaries were observed. Histological examination showed apparent differences between these two kinds of ovaries, whereas among male shrimp, there were no such differences. Contribution No. 3465 from the Institute of Oceanology. Chinese Academy of Sciences. The study was supported by the International Foundation for Science, Asian Fisheries Society and Chinese Climbing Project PDB 6–2, and Chinese National 863 Project (No. 863-819-01-05).     

Temperature effect on the life history of three types of Brachionus calyciflorus females

The effect of temperature on the life history characteristics of amictic females (AF), unfertilized mictic females (UMF) and fertilized mictic females (FMF) inBrachionus calyciflorus was studied with replicated individual cultures at 20°C, 25°C and 30°C, and with algaeScenedesmus obliquus for their food. There were highly significant effects of both temperature and female type, independently and interactively, on the duration of juvenile and post-reproduction periods, and the number of eggs produced by the rotifer per life cycle. Among all the temperature-female type combinations, all the juvenile periods of FMF, and the post-reproduction periods of UMF and FMF at 20°C, were the longest, and the number of eggs produced by an UMF at 30°C was the highest. There were highly significant effects of both temperature and female type on the duration of the reproduction period, but no clear correlation was observed between temperature and female type. The reproduction period of AF was longer than that of UMF and FMF. Only temperature influenced significantly the mean life-span of the three types of female. The duration of juvenile, reproduction and post-reproduction periods as well as the life-span of the three types of females were all reduced very significantly with rise of temperature, but the rates of reduction varied with female type. Among the three types of females, the number of eggs produced per life cycle by an UMF was the highest, and that of a FMF was the lowest. A significant relationship between the number of eggs produced per life cycle and temperature was observed only in the UMF. Projects supported by NSFC (No. 39870158), Natural Science Foundation of Anhui Province (No. 100330: No. 042416) and the Excellent Youth Foundation (No. 04043050).     

Chromosome behavior of heat shock induced triploid in Fenneropenaeus chinensis

Triploidy was induced in Chinese shrimpFenneropenaeus chinensis by 30±0.5°C heat shock treatment (initiated at 20 min after fertilization) for 10 min to inhibit the release of PB2 at 18.0°C. The highest triploid rate obtained was 84.5% in nauplius stage. The effect of heat shock treatment on meiosis and cleavage of eggs was investigated in this work aimed to establish efficient procedures for triploid induction and to gain understanding of the mechanism of triploid production. Three pronuclei that could be observed in the treated eggs under fluorescence microscope developed into triploid embryos. Some abnormal chromosome behavior was observed in heat shocked eggs. Contribution No. 4407 from the Institute of Oceanology, Chinese Academy of Sciences. This work was supported by Project under Major State Basic Development Program (G1999012009) and Knowledge Innovation Program of the Chinese Academy of Sciences (KZCX 2-211)     

STUDY ON EMBRYONIC DEVELOPMENT AND EARLY GROWTH OF TRIPLOID AND GYNOGENETIC DIPLOID LEFI—EYED FLOUNDER, PARALICHTHYS OLIVACEUS （T. et S. ）

The early effects of chromosomal manipulation of eggs and sperm on the yields of trip-loid and gynogenefic diploid larvae of Paralichthys olivaceus were investigated. Triploidy was achieved by cold shocking fen.ilized eggs at 0 - 2℃ for 45 minutes duration 5 minutes after fen.ilization, and the in-duced triploidy rates were 31.2% - 50% and the relative hatching rates were 53.3% - 99%. Gynnge-aetic diploids were obtained when eggs were inseminated with irradiated sperm and cold shocked at 0 -2℃ for 45 minutes duration 5 minutes after fertihzation. The induced gynogenetic diploid rates and the relative hatching rates were 94 % - 96 % and 48.5 % - 68.5 % respectively. The embryonic development of the triploid experimental group and of the gynogenetic diploid experimental group was delayed at first compared with the control group. But from the gastrula stage, it was not delayed anymore. There were no significant differences in the growth of the triploid experimental group larvae and the control group larvae, and in the growth of the gynogenetic diploid experimental group larvae and the control group larvae ac-cording to Student‘s t-test (α = 0.05). The relationship between the early growth of the triploid experi-mental group larvae and that of gynogenetic diploid experimental group larvae was also studied.     

Studies on extraction procedure and antioxidative activity of phlorotannins fromSargassum kjellmanianum

Known only in phaeophyceae, phlorotannins (brown algal polyphenols) are natural products with potential uses in pharmacology. This study yielded an extraction procedure to obtain high purity, high molecular weight phlorotannins fromSargassum kjellmanianum and revealed the characteristics of their infrared and flourescence spectra. The antioxidative activity of phlorotannins, which was about 2.6 times as strong as that of 0.02% BHT (tert-butyl-4-hydroxytoluene), showed potential for preventing oil rancidification. Contribution No. 2918 from the Institute of Oceanology, Chinese Academy of Sciences. The project was supported by The State Ninth-five Key Program of China. (No. 96-916-04-01).     

Effect of ration, temperature and body weight on growth of black porgy,Sparus macrocephalus (B.)

Growth of black porgy,Sparus macrocephalus, was studied experimentally at four ration levels from starvation to satiation and four temperatures ranging from 14.8 to 26.8°C. At maximum rations, the specific growth rate (SGR) increased with temperature and was a negative linear function of body weight. The relationship between SGR and ration levels was a decelerating curve. Stepwise regression was used to develop the predictive models for the specific growth rates. Maintenance rations and optimum rations both increased with increased temperature. The relationship between maintenance, optimum rations and body weight was influenced by temperature. Conversion efficiencies increased with ration from zero at maintenance ration to a peak at the optimum ration, then declined with further increased ration. Contribution No. 3127 from the Institute of Oceanology, Chinese Academy of Sciences. Project 39370542 supported by NSFC and study also supported by National Key Laboratory in the Institute of Hydrobiology, Chinese Academy of Sciences.     

PHYLOGENETIC RELATIONSHIPS AMONG PUFFERFISH OF GENUS TAKIFUGU BY RAPD ANALYSIS

AmpLifications with 300 10-base primers under predetermined optimal reaction yielded 2762 reproducible amplified fragments ranging from 200 to 3000 bp. Genetic distances among five species of Takifugu and Lagocephalus spadiceus as outgreup were calculated based on the presence/absence of the amplified fragments. Topological phylogenie trees of the 5 species of Takgfugu and the outgroup were gen-erated by Parsimony and Neighbor-Joining analysis based on RAPD data set. The genetic distance between T. rubripes and T. pseudommus was almost the same as that between individuals within each species, but was much smaller than that among T. rubripes, T. pseudommus and the other species. The molecu-lar data from RAPD analysis convincingly showed that T. rubripes and T. pseudommus should be the same species.