Molecular dissection of mercury-responsive transcriptome and sense/antisense genes in Medicago truncatula

We described a newly developed approach, namely next-generation tag sequencing, to identify global gene transcripts and complexity regulated by heavy metals in Medicago truncatula. Two cDNA libraries were generated from M. truncatula seedlings: treated and non-treated with the toxic heavy metal mercury Hg(II). With the large number of read-mapped genes generated, we observed that most of the genes were differentially expressed between the two libraries. In addition, several classes of new transcripts including transcription factors, antisense transcripts, and stress responsive genes were detected. The forty genes most altered in expression levels were associated with tolerance to environmental stress and secondary metabolism. Validation of genes by quantitative RT-PCR confirmed the results from deep-sequencing. Most of genes coding for metal transporters, sulfate metabolism, and cell wall solidification were significantly altered by Hg exposure. We also examined altered expression ratios of sense and antisense (S-AS) transcripts between the two libraries. By analyzing strand-specific information of read sequences, S-AS transcripts were found to be enriched with metal treatment. The transcriptome sequences were analyzed further with Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) and showed diverse biological functions and metabolic pathways under the metal stress.