Functional characterisation of Target of Rapamycin (TOR) signalling in Physcomitrella

TL;DR

This study explores how TOR signaling affects growth and development in the moss Physcomitrella, revealing its role in chloroplast function, protein synthesis, and cell cycle.

Contribution

The study functionally characterizes TOR signaling in the non-vascular plant Physcomitrella, identifying key components and effects of TOR inhibition.

Findings

TOR inhibition in Physcomitrella delays cell-cycle progression and development.

Rapamycin and AZD8055 induce chlorosis and inhibit photosynthesis in Physcomitrella.

PpLST8 can substitute its yeast homolog to support cell growth.

Abstract

The evolutionary conserved TOR kinase positively controls growth and development of the moss Physcomitrella, development and function of its chloroplasts, its protein synthesis, and cell-cycle progression.

Target of Rapamycin (TOR) is a conserved protein kinase and an important signalling hub in eukaryotes. The moss Physcomitrella (Physcomitrium patens) is a model organism for plant physiology, development, and evolution. However, little is known about TOR signalling in non-vascular plants, including Physcomitrella. Here, we report the effects of inhibiting TOR signalling in Physcomitrella. We identified and characterised Physcomitrella 12-kDa FK506-binding protein (FKBP12), which binds TOR in the presence of rapamycin. Whereas the growth of wild-type protonema is unaffected by rapamycin, overexpressing endogenous FKBP12 rendered the plant susceptible to the inhibitor in a dose-dependent manner. Likewise, protonema growth was inhibited when the TOR-specific inhibitor AZD8055 was present in the culture. We show that rapamycin and AZD8055 have pleiotropic effects, as they delay cell-cycle progression and development, induce chlorosis, inhibit photosynthesis, and alter total protein content. Additionally, we identified and characterised PpTOR, PpLST8, and PpRAPTOR, key components of TOR complex 1 (TORC1), whereas RICTOR and mSIN1 of TORC2 are not encoded by the P. patens genome. We found that PpLST8 substitutes its homolog in yeast to allow cell growth. Physcomitrella mutants were generated with a conditional downregulation of PpTOR, PpLST8, and PpRAPTOR, respectively. They were impaired in growth. Finally, we show TOR-dependent phosphorylation of a well-known TOR phospho-target, the ribosomal protein RPS6, via LC–MS/MS. Collectively, our results show that Physcomitrella growth and development is positively controlled by a conserved TOR kinase. We suggest to further dissect TOR signalling in Physcomitrella in order to elucidate signalling integration via TORC1 in plants.

The online version contains supplementary material available at 10.1007/s00299-026-03722-y.

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