Biology subjects

Ferreras-Garrucho, G.

Publications and source records attributed to Ferreras-Garrucho, G..

3 recordsLinked to original sources

bioRxiv2026

Gain-of-function CCaMK in rice overrides genetic and anatomical barriers to arbuscular mycorrhizal colonisation

Arbuscular mycorrhizal (AM) symbiosis is conserved across land plants and is the default nutrient uptake strategy in nature. Within roots, AM colonisation is tightly patterned and dynamically tuned by nutritional cues. Multiple genetic modules contribute to this regulation, including the phosphate starvation response, DWARF14-LIKE (D14L) karrikin signalling, and the common symbiosis signalling pathway (CSSP). Transcriptional overlap among these has led to the hypothesis that phosphate starvation and D14L signalling act upstream of the CSSP. Here, we examined the epistatic relationship between D14L and CSSP in rice. Overexpression of an autoactive gain-of-function CCaMK (gofCCaMKox) restored AM colonisation and symbiosis marker gene expression in d14l mutants to wild-type levels or above, whereas overexpression of wild-type CCaMK did not, confirming that CSSP operates downstream of D14L signalling. However, gofCCaMKox did not rescue the d14l mesocotyl elongation phenotype, supporting a bifurcation of D14L into developmental and symbiotic outputs. Unexpectedly, gofCCaMKox also expanded fungal access to normally restrictive tissue domains (the meristematic zone and endodermis) assigning a role for CCaMK activation in defining root zone and cell-type competence for AM colonisation. Despite restored colonisation, introduction of gofCCaMKox into d14l produced arbuscules, which however were less developed and had increased hyphal septation, revealing a CCaMK-independent role for D14L in intraradical colonisation and arbuscule development. Transcriptome profiling resolved AM-relevant genes into modules controlled by CCaMK activation alone, in combination with D14L, or requiring additional colonisation-associated cues, and further suggested CCaMK primarily acts through AP2 transcription factors. Together, these findings reinforce CCaMK as a master regulator of AM symbiosis at the genetic, transcriptomic and anatomical levels while uncovering CCaMK-independent functions of D14L in arbuscule development.

plant biology↗

bioRxiv2025

Spatiotemporal regulation of arbuscular mycorrhizal symbiosis at cellular resolution

Arbuscular mycorrhizal (AM) symbiosis develops through successive colonization of root epidermal and cortical cells, culminating in the formation of arbuscules, tree-like intracellular structures that are transient yet essential sites of nutrient exchange. To dissect the cellular and structural complexity of AM establishment in rice roots colonized by Rhizophagus irregularis, we applied dual-species spatial transcriptomics to simultaneously monitor plant and fungal gene transcripts at single-cell resolution. This approach revealed surprising differences in transcriptional activity between fungal structures and showed that morphologically similar arbuscules can be transcriptionally distinct. These findings suggest hidden functional diversity among arbuscules at single-cell resolution. Because arbuscules form and degenerate within only a few days, we further sought to capture translational activities across their life span. We pioneered AM-inducible TRAP-seq (Translating Ribosome Affinity Purification followed by RNA-seq) using stage-specific promoters, enabling cell-type- and stage-resolved profiling in AM symbiosis. This revealed extensive spatiotemporal reprogramming of nutrient transport and signalling, with distinct sets of phosphate, nitrogen, and carbon transporters and regulators induced or repressed at different stages of arbuscule development, suggesting that nutrient exchange is dynamically regulated across the arbuscule life cycle. More broadly, cell wall biosynthesis genes and key defence markers were suppressed during arbuscule formation, whereas at a later stage, defence markers were strongly upregulated, suggesting a host-driven shift towards arbuscule termination. Together, these findings highlight the nuanced and dynamic regulation of AM symbiosis at the cellular level, refining our understanding of how nutrient exchange and fungal development are coordinated in space and time.

plant biology↗

bioRxiv2025

Defining the pre-symbiotic transcriptional landscape of rice roots

Plants interact with a plethora of organisms in the rhizosphere, with outcomes that range from detrimental to beneficial. Arbuscular mycorrhizal (AM) symbiosis is the most ubiquitous beneficial plant interaction in terrestrial ecosystems and involves soil borne fungi of the Glomeromycotina. It is believed that plants detect diagnostic signals for the discrimination between beneficial arbuscular mycorrhizal (AM) fungi and parasitic fungi during the pre-symbiotic molecular crosstalk. Here, we investigated the transcriptome of rice roots upon exposure to the complete cocktail of fungal exudates from either beneficial Rhizophagus irregularis or pathogenic Magnaporthe oryzae. We report that regardless of the exudate donor species, the transcriptional response lacked diagnostic differences. Instead, the profiles were marked by the common suppression of symbiosis signalling components, accompanied by the induction of a generic stress response (GSR) and defense-related signature, which was retained in a suite of symbiosis signalling mutants impaired at different stages of symbiosis development. However, upon permitting physical engagement with AM fungi, a striking reversion in the transcriptional responses occurred marked by the simultaneous relaxation of symbiosis signalling suppression and down-regulation of defense-related and GSR markers, overall comparable between wild-type and mutants. Our data therefore reveal that rather than specific recognition in the rhizosphere, a sequence of signals orchestrates stress, immunity and symbiosis, pivoting towards symbiosis potentially at the stage of plant-fungal contact formation. Significance StatementMost plant roots engage in symbiosis with beneficial mycorrhizal fungi. Before physical contact, early signal exchanges involve diffusible molecules. The bouquet of chemicals released by beneficial versus detrimental fungi has been proposed to convey specificity for signal transduction, transcriptional output and responses. Here we characterised the transcriptomes of rice roots to exudates from beneficial and pathogenic fungi, and found that they lack specific signatures but elicit a generic stress and immune response and suppression of symbiosis signalling. This profile was reversed, along with activation of symbiotic transcriptional responses, only when AM fungi were allowed to establish physical contact. This reveals the need for additional, unknown, layers of signal exchanges for plants to commit to symbiosis.

plant biology↗