Mn²⁺ modulates the expression of cellulase genes in Trichoderma reesei Rut-C30 via calcium signaling

The filamentous fungus Trichoderma reesei Rut-C30 is one of the most vital fungi for the production of cellulases, which can be used for biofuel production from lignocellulose. Nevertheless, the mechanism of transmission of external stimuli and signals in modulating cellulase production in T. reesei Rut-C30 remains unclear. Calcium is a known second messenger regulating cellulase gene expression in T. reesei.

In this study, we found that a biologically relevant extracellular Mn²⁺ concentration markedly stimulates cellulase production, total protein secretion, and the intracellular Mn²⁺ concentration of Rut-C30, a cellulase hyper-producing strain of T. reesei. Furthermore, we identified two Mn²⁺ transport proteins, designated as TPHO84-1 and TPHO84-2, indicating that they are upstream in the signaling pathway that leads to cellulase upregulation. We also found that Mn²⁺ induced a significant increase in cytosolic Ca²⁺ concentration, and that this increased cytosolic Ca²⁺ might be a key step in the Mn²⁺-mediated regulation of cellulase gene transcription and production. The utilization of LaCl₃ to block plasma membrane Ca²⁺ channels, and deletion of crz1 (calcineurin-responsive zinc finger transcription factor 1) to interrupt calcium signaling, showed that Mn²⁺ exerts the induction of cellulase genes via calcium channels and calcium signaling. To substantiate this, we identified a Ca²⁺/Mn²⁺ P-type ATPase, TPMR1, which could play a pivotal role in Ca²⁺/Mn²⁺ homeostasis and Mn²⁺ induction of cellulase genes in T. reesei Rut-C30.

Taken together, our results revealed for the first time that Mn²⁺ stimulates cellulase production, and demonstrates that Mn²⁺ upregulates cellulase genes via calcium channels and calcium signaling. Our research also provides a direction to facilitate enhanced cellulase production by T. reesei.