Salas, E. | bioRxiv Science

bioRxiv2026

Large-scale forward genetics reveals genetically separable components of plant cell proliferative capacity

Plant cell cultures underpin regeneration, transformation and emerging applications in biotechnology and plant cell agriculture, yet the genetic determinants of proliferative performance remain poorly characterized. Here, we combined large-scale forward genetics with automated image-based phenotyping to quantify 27,648 calli representing 2,880 Arabidopsis thaliana T-DNA insertion lines. Of these, 2,456 lines produced calli smaller than the WT population mean, whereas only 190 produced larger calli, revealing a strongly asymmetric phenotypic distribution. Enhanced callus growth was not consistently associated with increased vegetative growth, indicating that these traits can be genetically uncoupled. Validation of selected high-growth mutants in suspension culture identified lines with enhanced biomass accumulation, while serial propagation revealed independent variation in sustained proliferative capacity. Thus, callus growth, whole-plant growth, short-term suspension biomass accumulation and sustained proliferation represent distinct, not necessarily coupled components of proliferative performance. These findings establish a scalable framework for identifying genetic determinants relevant to the development of improved plant cell production lines.

plant biology↗

bioRxiv2025

Branched-chain amino acid assimilation promotes mixotrophy of ammonia-oxidizing archaeal sponge symbionts

Ammonia-oxidizing archaea (AOA) frequently form symbiotic associations with marine sponges. While free-living AOA are generally considered metabolically constrained chemolithoautotrophs, sponge-associated AOA encode for a branched-chain amino acid (BCAA) transporter, suggesting mixotrophic potential. Here, we test the unusual mixotrophic lifestyle of sponge-associated AOA by tracing the assimilation of 13C- and 15N-labeled BCAA in the sponge holobiont Ianthella basta. We demonstrate that BCAA degradation fuels ammonia oxidation and quantify BCAA uptake at the single-cell level by combining stable isotope probing, catalyzed reporter deposition fluorescence in situ hybridization, and nanoscale secondary ion mass spectrometry. Our results reveal that sponge-associated AOA are mixotrophic, assimilating BCAA as an additional carbon and nitrogen source. This metabolic adaptation may modulate BCAA availability in the holobiont, potentially regulating the hosts mTOR pathway. Collectively, our study reveals a novel nutritional interaction in sponge holobionts and challenges the perception of constrained metabolic capacities of AOA.

microbiology↗