Root systems play an essential role in ensuring plant productivity. Experiments conducted in controlled environments and simulation models suggest that root geometry and responses of root architecture to environmental factors should be studied as a priority. However, compared with aboveground plant organs, roots are not easily accessible by non-invasive analyses and field research is still based almost completely on manual, destructive methods. Contributing to reducing the gap between laboratory and field experiments, we present a novel phenotyping system (GROWSCREEN-Rhizo), which is capable of automatically imaging roots and shoots of plants grown in soil-filled rhizotrons (up to a volume of ~18 L) with a throughput of 60 rhizotrons per hour. Analysis of plants grown in this setup is restricted to a certain plant size (up to a shoot height of 80 cm and root-system depth of 90 cm). We performed validation experiments using six different species and for barley and maize, we studied the effect of moderate soil compaction, which is a relevant factor in the field. First, we found that the portion of root systems that is visible through the rhizotrons’ transparent plate is representative of the total root system. The percentage of visible roots decreases with increasing average root diameter of the plant species studied and depends, to some extent, on environmental conditions. Second, we could measure relatively minor changes in root-system architecture induced by a moderate increase in soil compaction. Taken together, these findings demonstrate the good potential of this methodology to characterise root geometry and temporal growth responses with relatively high spatial accuracy and resolution for both monocotyledonous and dicotyledonous species. Our prototype will allow the design of high-throughput screening methodologies simulating environmental scenarios that are relevant in the field and will support breeding efforts towards improved resource use efficiency and stability of crop yields.

New techniques and approaches have been developed for root phenotyping recently; however, rapid and repeatable non-invasive root phenotyping remains challenging. Here, we present GrowScreen-PaGe, a non-invasive, high-throughput phenotyping system (4 plants min-1) based on flat germination paper. GrowScreen-PaGe allows the acquisition of time series of the developing root systems of 500 plants, thereby enabling to quantify short-term variations in root system. The choice of germination paper was found to be crucial and paperroot interaction should be considered when comparing data from different studies on germination paper. The system is suitable for phenotyping dicot and monocot plant species. The potential of the system for high-throughput phenotyping was shown by investigating phenotypic diversity of root traits in a collection of 180 rapeseed accessions and of 52 barley genotypes grown under control and nutrient-starved conditions. Most traits showed a large variation linked to both genotype and treatment. In general, root length traits contributed more than shape and branching related traits in separating the genotypes. Overall, results showed that GrowScreen-PaGe will be a powerful resource to investigate root systems and root plasticity of large sets of plants and to explore the molecular and genetic root traits of various species including for crop improvement programs

Transient expression systems allow the rapid production of recombinant proteins in plants. Such systems can be scaled up to several hundred kilograms of biomass, making them suitable for the production of pharmaceutical proteins required at short notice, such as emergency vaccines. However, large‐scale transient expression requires the production of recombinant Agrobacterium tumefaciens strains with the capacity for efficient gene transfer to plant cells. The complex media often used for the cultivation of this species typically include animal‐derived ingredients that can contain human pathogens, thus conflicting with the requirements of good manufacturing practice (GMP). We replaced all the animal‐derived components in yeast extract broth (YEB) cultivation medium with soybean peptone, and then used a design‐of‐experiments approach to optimize the medium composition, increasing the biomass yield while maintaining high levels of transient expression in subsequent infiltration experiments. The resulting plant peptone Agrobacterium medium (PAM) achieved a two‐fold increase in OD600 compared to YEB medium during a 4‐L batch fermentation lasting 18 h. Furthermore, the yields of the monoclonal antibody 2G12 and the fluorescent protein DsRed were maintained when the cells were cultivated in PAM rather than YEB. We have thus demonstrated a simple, efficient and scalable method for medium optimization that reduces process time and costs. The final optimized medium for the cultivation of A. tumefaciens completely lacks animal‐derived components, thus facilitating the GMP‐compliant large‐scale transient expression of recombinant proteins in plants.

Root system architecture (RSA) is a target for breeding crops with effective nutrient and water use. Breeding can use populations designed to map quantitative trait loci (QTL). Here we non-invasively phenotype roots and leaves of the 16 foundation parents of two multi-parent advanced generation inter-cross (MAGIC) populations, covering diversity in spring (CSIRO MAGIC) and winter (NIAB MAGIC) wheats. RSA components varied after 16 days in the upgraded, paper-based imaging platform, GrowScreen-PaGe: lateral root length 2.2 fold; total root length, 1.9 fold; and seminal root angle 1.2 fold. RSA components total and lateral root length had the highest root heritabilities (H2) (H2 = 0.4 for CSIRO and NIAB parents) and good repeatability (r = 0.7) in the GrowScreen-PaGe. These can be combined with leaf length (H2 = 0.8 CSIRO; 0.7 NIAB) and number (H2 = 0.6 CSIRO; 0.7 NIAB) to identify root and shoot QTL to breed for wheats with vigorous RSA and shoot growth at establishment, a critical phase for crop productivity. Time resolved phenotyping of MAGIC wheats also revealed parents to cross in future for growth rate traits (fastest: Robigus–NIAB and AC Barrie–CSIRO; slowest Rialto–NIAB and G204 Xiaoyan54–CSIRO) and root: shoot allocation traits (fast growers grew roots, notably laterals, quicker than leaves, compared to slow growers).

In face of the alarming world population growth predictions and its threat to food security, the development of sustainable fertilizer alternatives is urgent. Moreover, fertilizer performance should be assessed not only in terms of yield but also root system development, as it impacts soil fertility and crop productivity. Fertilizers containing a polysulfide matrix (PS) with dispersed struvite (St) were studied for S and P nutrition due to their controlled-release behavior. Soybean cultivation with St/PS composites provided superior biomass compared to a reference of triple superphosphate (TSP) with ammonium sulfate (AS), with up to 3 and 10 times higher mass of shoots and roots, respectively. Additionally, St/PS achieved a 22% sulfur use efficiency against only 8% from TSP/AS. Root system architectural changes may explain these results, with higher proliferation of second order lateral roots in response to struvite ongoing P delivery. Overall, the composites showed great potential as efficient controlled-release fertilizers for enhanced soybean productivity.