Dr Kim Tran

An investigation was conducted over three estuaries in SE Australia with a gradient in metal(loid) contamination to assess metal(loid) (Cu, Zn, As, Se, Cd and Pb) accumulation and transport within the halophytic saltmarsh rush, Juncus kraussii. Sydney Olympic Park exhibited the most elevated metal(loid) contamination, followed by Hunter Wetlands and Lake Macquarie. J. kraussii exhibited a strong ability to restrict metal(loid) movement into the root system, with the exception of cadmium (BCFs < 1.0) and unrestricted flow from root to culm excepting Se, Cd (TFs < 1). Pb and Zn exhibited elevated translocation between roots and culms (TF 4.4 and 7.3, respectively). Despite barriers for uptake into the below-ground tissues, most metal(loid)s were accumulated to the roots with environmental dose (except for Cu and Cd) and linear relationships were present between the root and culm (for As and Se) and the sediment and culm (for As, Se, Cd, and Pb).

We examined the patterns of uptake and partitioning of metal(loid)s in Suaeda australis from three highly urbanised estuaries (Sydney Olympic Park, Hunter Wetlands and Lake Macquarie) in NSW, Australia. Of these, Sydney Olympic Park was found to be the most contaminated estuary in terms of combined sediment metal(loid) load, followed by Hunter Wetlands and lowest in Lake Macquarie (via PERMANOVA). Uptake in roots was greater for the essential metals Cu and Zn along with the non-essential metal Cd and the metalloid Se (root BCFs >1) and lower for Pb and As (root BCFs <1). Substantial barriers for translocation from roots to stems were identified for all metal(loid)s (stem TFs; 0.07¿0.68). Conversely, unrestricted flow from stems to leaves was observed for all metal(loid)s at unity or higher (leaf TFs = 1). Strong linear relationships between sediment and root for Zn and Pb were observed, indicating roots as a useful bioindicator.

Marine molluscs, such as oysters, respond to estrogenic compounds with the induction of the egg yolk protein precursor, vitellogenin (Vtg), availing a biomarker for estrogenic pollution. Despite this application, the precise molecular mechanism through which estrogens exert their action to induce molluscan vitellogenesis is unknown. As a first step to address this question, we cloned a gene encoding Vtg from the Sydney rock oyster Saccostrea glomerata (sgVtg). Using primers designed from a partial sgVtg cDNA sequence available in Genbank, a full-length sgVtg cDNA of 8498 bp was obtained by 5'- and 3'-RACE. The open reading frame (ORF) of sgVtg was determined to be 7980 bp, which is substantially longer than the orthologs of other oyster species. Its deduced protein sequence shares the highest homology at the N- and C-terminal regions with other molluscan Vtgs. The full-length genomic DNA sequence of sgVtg was obtained by genomic PCR and genome walking targeting the gene body and flanking regions, respectively. The genomic sequence spans 20 kb and consists of 30 exons and 29 introns. Computer analysis identified three closely spaced half-estrogen responsive elements (EREs) in the promoter region and a 210-bp CpG island 62 bp downstream of the transcription start site. Upregulation of sgVtg mRNA expression was observed in the ovaries following in vitro (explants) and in vivo (tank) exposure to 17ß-estradiol (E2). Notably, treatment with an estrogen receptor (ER) antagonist in vitro abolished the upregulation, suggesting a requirement for an estrogen-dependent receptor for transcriptional activation. DNA methylation of the 5' CpG island was analysed using bisulfite genomic sequencing of the in vivo exposed ovaries. The CpG island was found to be hypomethylated (with 0-3% methylcytosines) in both control and E2-exposed oysters. However, no significant differential methylation or any correlation between methylation and sgVtg expression levels was observed. Overall, the results support the possible involvement of an ERE-containing promoter and an estrogen-activated receptor in estrogen signalling in marine molluscs.