Lisheng Wu

In this study, the behavioral ethogram of Amphiprion clarkii during the growth phase prior to sexual differentiation was summarized based on behavioral observations in three social environments. These behaviors can be classified into four categories: in addition to normal behaviors, the other three categories of behaviors—threatening, agonistic, and appeasing behaviors—represent different intentions in interactions with other individuals. Subsequently, the personalities of each individual were assessed by testing their reactions to intruders. These individuals mainly exhibited two distinct personality types: bold-aggressive and shy-submissive. In pairing experiments, the interactive behaviors of the anemonefish were observed in pairing combinations of different body sizes and personalities. The impact of personality on the establishment of a stable social hierarchy was confirmed by significant differences in the success rates of different pairing combinations, with the frequency of appeasing behaviors being the main factor influencing the success rate. Our results suggested that in natural waters, when juvenile individuals migrate among host anemones, shy-submissive individuals are more likely to be accepted due to their appeasing behaviors towards larger individuals, thus avoiding the risk of being attacked and bitten, and benefiting the survival of the individual. Conversely, bold-aggressive individuals are more likely to be driven away to another host anemone due to their unwillingness to settle for a lower-ranked status, thereby contributing to population dispersal and increasing opportunities for gene exchange between populations.

In this study, we identified the important contribution of frontal bone remodeling in shaping the ‘sunken head and humpback’ appearance in C. altivelis. Our investigation identified a developmental milestone at a total length of 5–6 cm, making the onset of its morphologic specialization in this species. A comparative analysis with closely related species reveals heightened activity in the frontal osteoblasts of the humpback grouper, potentially providing a physiological basis for its remodeling. Furthermore, our findings highlight that a significant upregulation in the expression levels of Ihhb, Ptch1, and Gli2a genes was seen in C. altivelis within the specified developmental stage, indicating an important involvement of the Ihhb-Ptch1-Gli2a signaling pathway in initiating the morphological specialization. We hypothesized that Ihh signaling could be attributed to shifts in mechanical stress, resulting from muscle traction on the frontal bone due to changes in swimming patterns during development. This study not only offers significant insights into unraveling the molecular mechanisms that govern phenotypic specialization and ecological adaptations in the humpback grouper but also serves as a valuable reference for studies on fishes with a controversial morphology and molecular phylogeny.

Diel rhythms in feeding and spawning were investigated in Centropages tenuiremis from Xiamen Bay in March to May, 2006. Circular statistics were used to determine the peak time of spawning. The results showed that the feeding activities of females were stably higher at night-time, and there was a remarkable earlier shift in spawning peak time with warmer seawater. Thus, the lag times between peak times of gut pigment content and spawning were shortened with the increase of temperature. It suggested that there was a direct effect of feeding rhythms on egg production variations in copepods, and the seawater temperature would work on the converting time and then influence the spawning peak time. So the effect of temperature cannot be ignored in the investigation of the effects of feeding on egg production.

To study the molecular response mechanisms of copepod to nickel stress, a suppression subtractive hybridization (SSH) cDNA library of Pseudodiaptomous annandalei under nickel stress was constructed by using SSH technique, and a total of 140 clones were randomly picked from the growing colonies and identified by PCR. The recombinant rate of the library was 98.6%, and the volume of the library was 1.12 x 10(6) cfu. After the recombinant plasmids were sequenced, a partial cDNA fragment of ferritin was recognized based on BLAST searches in NCBI, with a size of 859 bp and continuously encoding 170 amino acid residues. The semi-quantitative PCR results showed that the ferritin cDNA under 24 h nickel stress was distinctly up-regulated. The successful construction of the SSH library and the obtaining of ferritin cDNA fragment would supply basis for the further study of the molecular response mechanisms of copepod to nickel stress.

A method of extracting DNA from individual copepod eggs was introduced for this study which included a modified proteinase K procedure and an efficient DNA sedimentation process. DNA was extracted from egg samples including freshly spawned eggs from the three copepod species Apocyclops borneoensis, Centropages tenuiremis and Calanus sinicus, together with 18 resting eggs separated from different sediment layers. A short fragment of the 28S rDNA (~300 bp) sequence that varied between copepod species was amplified and sequenced. These sequences were used to construct a UPGMA tree which helped to assess species composition and the distribution of copepods buried in the sediments. The results showed that C. tenuiremis and Acartia pacifica were closely clustered in the tree with egg samples from deeper sediment layers, whereas A. borneoensis and C. sinicus were grouped with surface egg samples. Species composition in the sediments varied between sediment layers and sampling locations. The DNA extraction method was valid for analysing individual copepod eggs with different egg-spawning types and sizes and the results helped us to reconstruct the copepod egg composition and distribution in the sediments. We believe that the technique has a wide usage in analysing the copepod egg bank in sediments, and possibly even for other zooplankton.