Project description:Illumina HiSeq technology was used to generate mRNA profiles from two strains of Trametes versicolor. Mycelium of Trametes versicolor BRFM1218 and Trametes versicolor 1956-1252 were harvested after 2 and 4 weeks of incubation on 4% malt agar medium and used for total RNA extraction. Paired-end reads of 100 bp were generated and aligned to Trametes versicolor (https://mycocosm.jgi.doe.gov/Trave1/Trave1.home.html) reference transcripts using CLC Genomics Workbench 7.5.1.
Project description:Trametes hirsuta AH28-2 is a white-rot basidiomycete originally isolated from rotting wood in China. This strain can secrete high levels of extracellular laccase induced by copper and o-toluidine with great potential applications in industry and environment. However, the proteomics and mechanisms for regulating laccases from T. hirsuta AH28-2 fungi at the protein level have not been investigated. To explore the functional factors related to laccases, label-free quantification proteomics was used to identify proteins produced by T. hirsuta AH28-2, and the differentially expressed proteins (DEPs) were compared in the control, 2 mM o-toluidine and 50 uM copper ion cultures of T. hirsuta AH28-2 for 48h. Here, across all samples, a total of 5,089 proteins were quantified, of which 504 exhibited significant differential expression (278 up-regulated and 226 down-regulated) in the o-toluidine induction group, and 447 with significant differential expression (282 up-regulated and 165 down-regulated) in the copper induction group. These DEPs from o-toluidine induction group were significantly enriched in the pathways of sphingolipid metabolism, toluene degradation, DNA replication and drug metabolism-cytochrome P450, and DEPs from copper induction group were significantly enriched in aminobenzoate degradation and ABC transporters pathways. Furthermore, a total of 120 and 119 DEPs identified in o-toluidine- and copper- induction group were predicted as TFs, respectively. Protein-protein interaction networks analyses showed that Zn2Cys6 (GME7257_g and GME6689_g) transcription factors possessed strong protein interaction with laccases in o-toluidine- and copper-induction group. Several proteins with different expression were consistent with the proteomics of T. hirsuta AH28-2 by parallel reaction monitoring (PRM) verification.
