ABSTRACT
High amount of copper is toxic to most organisms, but endophytic fungi can develop survival strategies to tolerate and respond to environmental stressors such as heavy metal contaminants. While high copper induces oxidative stress, it is still unclear which genes are associated with copper tolerance. Here, we performed a metatranscriptome analysis of endophytic fungi isolated from a black nightshade plant Solanum nigrum L. growing on mine tailings of a gold processing area. Initial screening revealed the presence of a copper-tolerant strain of Fusarium oxysporum, designated as IB-SN1W, which tolerated up to 1000 ppm and 300 ppm copper in solid and liquid media, respectively. Differential gene expression analysis by RNA sequencing showed that 23% of contigs are uniquely expressed in the copper-treated fungus. These genes are involved in copper ion import, polyamine transport, oxidoreductase activity, and oxidative stress response. Catalase transcripts were also highly upregulated in IB-SN1W compared to a non-tolerant F. oxysporum strain. Catalase inhibition decreased copper-tolerance in IB-SN1W, while the addition of antioxidants prevented the copper-dependent growth inhibition in the non-tolerant strain. Overall, these results suggest that oxidative stress response contributes to copper tolerance in F. oxysporum.
Subject(s)
Copper , Fusarium , Oxidative Stress , Transcriptome , Copper/pharmacology , Fusarium/drug effects , Fusarium/genetics , Gene Expression Profiling , Genes, Fungal/geneticsABSTRACT
Milkfish (Chanos chanos Forsskal) is an important aquaculture product and is the sole extant species of the family Chanidae (order Gonorynchiformes). While there are already several reports regarding milkfish aquaculture, studies on milkfish immunity and gene expression are very limited. In this study, we showed that Aeromonas hydrophila induces inflammation in milkfish liver. We identified a milkfish C-type lectin-like domain containing proteins (CTLDcps) gene, designated as CcClec, which was upregulated in respond to A. hydrophila. Full-length sequencing was performed using Rapid Amplification of cDNA Ends (RACE PCR) to produce a complete Coding DNA Sequence (CDS) of the gene. The CcClec gene encoded a predicted protein of 340 amino acids containing two CTLDs that may potentially bind carbohydrates, especially sucrose and cellobiose. The CcClec mRNA transcript was expressed highest in the liver, followed by head kidney, brain, heart, gills, spleen, and midgut. CcClec transcripts were upregulated in damaged liver upon exposure to A. hydrophila. Overall, these findings demonstrated that CcClec is implicated in milkfish innate immunity, and is most highly expressed in the liver, suggesting a role of the liver in the milkfish immune system.