Causes and consequences of experimental variation in Nicotiana benthamiana transient expression
Authors: Tang, S. N., Szarzanowicz, M., Lanctot, A., Sirirungruang, S., Kirkpatrick, L. D., Drako, K., Alamos, S., Cheng, L., Waldburger, L. M., Liu, S., Huang, L., Akyuz Turumtay, E., Kazaz, S., Baidoo, E., Eudes, A., Thompson, M., Shih, P.
The study systematically examines sources of variability in Agrobacterium tumefaciens-mediated transient expression in Nicotiana benthamiana, analyzing a large dataset of 1,915 plants collected over three years. It demonstrates that normalization methods must be validated for each experimental context and provides a statistical model and power analysis framework to determine appropriate sample sizes for detecting specific effect sizes, offering practical guidelines to improve reproducibility in quantitative plant and synthetic biology studies.
The study investigated how barley (Hordeum vulgare) adjusts mitochondrial respiration under salinity stress using physiological, biochemical, metabolomic and proteomic approaches. Salt treatment increased respiration and activated the canonical TCA cycle, while the GABA shunt remained largely inactive, contrasting with wheat responses.
The study investigates the wheat Pm3 NLR allelic series, revealing that near-identical Pm3d and Pm3e alleles confer broad-spectrum resistance by recognizing multiple, structurally diverse powdery mildew effectors. Using chimeric NLR constructs, the authors pinpoint specificity-determining polymorphisms and demonstrate that engineered combinations of Pm3d and Pm3e further expand effector recognition, showcasing the potential for durable wheat protection through NLR engineering.
The study generated deep proteome and phosphoproteome datasets from guard cell‑enriched tissue to examine how phosphorylation regulates stomatal movements. Comparative analysis revealed increased phosphorylation of endomembrane trafficking and vacuolar proteins in closed stomata, supporting a role for phospho‑regulated trafficking in stomatal dynamics.
Uncovering the Molecular Regulation of Seed Development and Germination in Endangered Legume Paubrasilia echinata Through Proteomic and Polyamine Analyses
Authors: Vettorazzi, R. G., Carrari-Santos, R., Sousa, K. R., Oliveira, T. R., Grativol, C., Olimpio, G., Venancio, T. M., Pinto, V. B., Quintanilha-Peixoto, G., Silveira, V., Santa-Catarna, C.
The study examined seed maturation and germination in the endangered legume Paubrasilia echinata using proteomic and polyamine analyses at 4, 6, and 8 weeks post-anthesis, identifying over 2,000 proteins and linking specific polyamines to developmental stages. Mature seeds (6 weeks) showed elevated proteasome components, translation machinery, LEA proteins, and heat shock proteins, while polyamine dynamics revealed putrescine dominance in early development and spermidine/spermine association with desiccation tolerance and germination. These findings uncover dynamic molecular shifts underlying seed development and provide insights for conservation and propagation.
Using CRISPR‑Cas9, researchers knocked down CCD7 or CCD8 in Nicotiana benthamiana to suppress strigolactone synthesis, producing compact plants with a 45%–50% smaller spatial footprint while preserving recombinant protein yields (GFP and rituximab). The mutants showed altered leaf proteome, auxin/cytokinin balance, and metabolic fluxes without affecting overall growth rate, demonstrating suitability for indoor vertical farming biopharma production.
Polyphenol oxidase mutant Nicotiana benthamiana plants increase yield and purity of recombinant proteins and enable studies of proteins in their native state.
The authors created two Nicotiana benthamiana lines with CRISPR-mediated knockouts of two polyphenol oxidase genes, which exhibited slightly accelerated growth and retained normal transient GFP expression. These ppo-deficient plants produced leaf extracts that remained greener with markedly reduced enzymatic browning and protein crosslinking, leading to a nearly fourfold increase in yield and improved purity of a transiently expressed His‑tagged tomato protease. The study demonstrates that PPO depletion can enhance recombinant protein recovery from plant tissue.
The study provides a comprehensive proteomic analysis of seed mitochondria from white lupin, revealing fully assembled OXPHOS complexes ready for immediate energy production upon imbibition. Quantitative mass‑spectrometry identified 1,162 mitochondrial proteins, highlighting tissue‑specific transporter and dehydrogenase profiles and dynamic remodeling during early germination, while many uncharacterized proteins suggest novel legume‑specific functions.
Regenerative agriculture effects on biomass, drought resilience and 14C-photosynthate allocation in wheat drilled into ley compared to disc or ploughed arable soil
Authors: Austen, N., Short, E., Tille, S., Johnson, I., Summers, R., Cameron, D. D., Leake, J. R.
Regenerative agriculture using a grass-clover ley increased wheat yields and macroaggregate stability despite reduced root biomass, but did not enhance soil carbon sequestration as measured by 14C retention. Drought further decreased photosynthate allocation to roots, especially in ley soils, while genotype effects on yield were minimal.
Light on its feet: Acclimation to high and low diurnal light is flexible in Chlamydomonas reinhardtii
Authors: Dupuis, S., Chastain, J. L., Han, G., Zhong, V., Gallaher, S. D., Nicora, C. D., Purvine, S. O., Lipton, M. S., Niyogi, K. K., Iwai, M., Merchant, S. S.
The study examined how prior light‑acclimation influences the fitness and rapid photoprotective reprogramming of Chlamydomonas during transitions between low and high diurnal light intensities. While high‑light‑acclimated cells struggled to grow and complete the cell cycle after shifting to low light, low‑light‑acclimated cells quickly remodeled thylakoid ultrastructure, enhanced photoprotective quenching, and altered photosystem protein levels, recovering chloroplast function within a single day. Transcriptomic and proteomic profiling revealed swift induction of stress‑response genes, indicating high flexibility in diurnal light acclimation.