Toxicity of BDE-47, BDE-99 and BDE-153 on swimming behavior of the unicellular marine microalgae Platymonas subcordiformis and implications for seawater quality assessment.

Polybrominated diphenyl ethers (PBDEs), a class of brominated flame retardants, have been extensively applied and eventually leached into the surrounding environment. Marine microalgae are not only the dominant primary producers of marine ecosystem, but also food source for aquaculture. PBDEs have been found to remarkably inhibit growth, photosynthesis and metabolism of marine microalgae. However, whether they also affect swimming behavior of marine motile microalgae remains unknown. We chose BDE-47, BDE-99 and BDE-153 as model PBDEs, and the unicellular marine green flagellate, Platymonas subcordiformis, as test organism to figure out this issue. After two-hour exposure, motile cells proportion (MOT), swimming velocity (VCL, VAP and VSL), and swimming pattern (LIN and STR) of P. subcordiformis were measured via computer assisted cell movement tracking. Results suggest that the three PBDEs not only reduced motile cells proportion and swimming velocity, but also altered swimming pattern. BDE-47 was more toxic than BDE-99, followed by BDE-153, indicating their toxicity decreased as bromination degree increases. Swimming ability of P. subcordiformis was even completely arrested when BDE-47 and BDE-99 at 32 µg/L. The impairment of swimming ability by PBDEs might thereby hinder growth and survival of marine microalgae, and subsequently threaten marine ecosystem and aquaculture industry. More importantly, this study implies that marine microalgae swimming behavior test is more efficiency and sensitive than traditional marine microalgal bioassays, like growth and photosynthesis tests. We suggest that although future work is needed, swimming behavior analysis of P. subcordiformis with MOT, VCL and VAP as endpoints can be developed as a low-cost, convenient, fast, reliable and sensitive method for seawater quality assessment.

Transcriptome analysis of sex-related genes in the blood clam Tegillarca granosa.

BACKGROUND: Blood clams (Tegillarca granosa) are one of the most commercial shellfish in China and South Asia with wide distribution in Indo-Pacific tropical to temperate estuaries. However, recent data indicate a decline in the germplasm of this species. Furthermore, the molecular mechanisms underpinning reproductive regulation remain unclear and information regarding genetic diversity is limited. Understanding the reproductive biology of shellfish is important in interpreting their embryology development, reproduction and population structure. Transcriptome sequencing (RNA-seq) rapidly obtains genetic sequence information from almost all transcripts of a particular tissue and currently represents the most prevalent and effective method for constructing genetic expression profiles. RESULTS: Non-reference RNA-seq, an Illumina HiSeq2500 Solexa system, and de novo assembly were used to construct a gonadal expression profile of the blood clam. A total of 63.75 Gb of clean data, with at least 89.46% of Quality30 (Q30), were generated which was then combined into 214,440 transcripts and 125,673 unigenes with a mean length of 1,122.63 and 781.30 base pairs (bp). In total, 27,325 genes were annotated by comparison with public databases. Of these, 2,140 and 2,070 differentially expressed genes (DEGs) were obtained (T05 T08 vs T01 T02 T04, T06 T07 vs T01 T02 T04; in which T01-T04 and T05-T08 represent biological replicates of individual female and male clams, respectively) and classified into two groups according to the evaluation of biological replicates. Then 35 DEGs and 5 sex-related unigenes, in other similar species, were investigated using qRT-PCR, the results of which were confirmed to data arising from RNA-seq. Among the DEGs, sex-related genes were identified, including forkhead box L2 (Foxl2), sex determining region Y-box (Sox), beta-catenin (ß-catenin), chromobox homolog (CBX) and Sex-lethal (Sxl). In addition, 6,283 simple sequence repeats (SSRs) and 614,710 single nucleotide polymorphisms (SNPs) were identified from the RNA-seq results. CONCLUSIONS: This study provided the first complete gonadal transcriptome data for the blood clam and allowed us to search many aspects of gene sequence information, not limited to gender. This data will improve our understanding of the transcriptomics and reproductive biology of the blood clam. Furthermore, molecular markers such as SSRs and SNPs will be useful in the analysis of genetic evolution, bulked segregant analysis (BSA) and genome-wide association studies (GWAS). Our transcriptome data will therefore provide important genetic information for the breeding and conservation of germplasm.