Comparative transcriptome analysis of ectomycorrhizal fungus Pisolithus albus in response to individual and combined stress of copper and cadmium.

An ectomycorrhizal fungus Pisolithus albus establishes the natural symbiosis with plant roots on extreme heavy metal (HM)-rich soil and enables their survival in toxic metal concentrations. Understanding P. albus key genes and pathways behind strong metal tolerance is crucial for its successful application in the rehabilitation of metal-contaminated barren lands. Therefore, this study aimed to analyze the whole transcriptome profile of P. albus under individual and combined metal stress of copper (Cu) and cadmium (Cd). At 480 µM Cu and 16 µM Cd toxic concentrations, P. albus has shown growth and survival and accumulated high metal (1.46 µg Cu and 1.13 µg Cd per mg of dry mycelia). The study found a stronger response of P. albus to single-metal stress in high concentration as compared to multi-metal stress in relatively lower concentration. Hence, the intensity of fungal response to HM stress is mainly determined by the metal concentration involved in stress. We have found a total of 11 pathways significantly associated with HM stress, among which amino acid, lipid, and carbohydrate metabolisms were highly affected. The functional enrichment of differentially expressed genes has shown the induced biosynthesis of arginine, melanin, metal chelating agents, membrane phospholipids, fatty acids, folate, pantothenate, ergothioneine, and other antioxidant agents; upregulation of zinc ion uptake, potassium transporters, and lysine degradation; and reduction of phosphatidylcholine degradation, incorrect protein folding, iron uptake, and potassium efflux as the top efficient tolerance mechanisms of P. albus against HM stress. The current study would contribute to understanding fungal HM tolerance and its further utilization in the bioremediation of metal-contaminated abandoned lands. The validation of RNA-sequencing analysis with RT-qPCR of selected genes showed the high credibility of the presented data.