A toolkit to rapidly modify root systems through single plant selection

Background The incorporation of root traits into elite germplasm is typically a slow process. Thus, innovative approaches are required to accelerate research and pre-breeding programs targeting root traits to improve yield stability in different environments and soil types. Marker-assisted selection (MAS) can help to speed up the process by selecting key genes or quantitative trait loci (QTL) associated with root traits. However, this approach is limited due to the complex genetic control of root traits and the limited number of well-characterised large effect QTL. Coupling MAS with phenotyping could increase the reliability of selection. Here we present a useful framework to rapidly modify root traits in elite germplasm. In this wheat exemplar, a single plant selection (SPS) approach combined three main elements: phenotypic selection (in this case for seminal root angle); MAS using KASP markers (targeting a root biomass QTL); and speed breeding to accelerate each cycle. Results To develop a SPS approach that integrates non-destructive screening for seminal root angle and root biomass, two initial experiments were conducted. Firstly, we demonstrated that transplanting wheat seedlings from clear pots (for seminal root angle assessment) into sand pots (for root biomass assessment) did not impact the ability to differentiate genotypes with high and low root biomass. Secondly, we demonstrated that visual scores for root biomass were correlated with root dry weight (r = 0.72), indicating that single plants could be evaluated for root biomass in a non-destructive manner. To highlight the potential of the approach, we applied SPS in a backcrossing program which integrated MAS and speed breeding for the purpose of rapidly modifying the root system of elite bread wheat line Borlaug100. Bi-directional selection for root angle in segregating generations successfully shifted the mean root angle by 30° in the subsequent generation (P ≤ 0.05). Within 18 months, BC2F4:F5 introgression lines were developed that displayed a full range of root configurations, while retaining similar above-ground traits to the recurrent parent. Notably, the seminal root angle displayed by introgression lines varied more than 30° compared to the recurrent parent, resulting in lines with both narrow and wide root angles, and high and low root biomass phenotypes. Conclusion The SPS approach enables researchers and plant breeders to rapidly manipulate root traits of future crop varieties, which could help improve productivity in the face of increasing environmental fluctuations. The newly developed elite wheat lines with modified root traits provide valuable materials to study the value of different root systems to support yield in different environments and soil types.

Root Morphology, Allometric Relations and Rhizosheath of Ancient and Modern Tetraploid Wheats (Triticum durum Desf.) in Response to Inoculation with Trichoderma harzianum T-22

Early root traits and allometrics of wheat are important for competition and use of resources. They are under-utilized in research and un-explored in many ancient wheats. This is especially true for the rhizosheath emerging from root-soil interactions. We investigated root morphology, root/shoot relations and the amount of rhizosheath of four tetrapoid wheat seedlings (30 days after emergence): the italian landrace Saragolle Lucana and modern varieties Creso, Simeto and Ciclope, and tested the hypothesis that inoculation with Trichoderma harzianum T-22 (T-22) enhances rhizosheath formation and affects wheat varieties differently. Overall growth of non-inoculated plants showed different patterns in wheat varieties, with Saragolle and Ciclope at the two extremes: Saragolle invests in shoot rather than root mass, and in the occupation of space with highest (p < 0.05) shoot height to the uppermost internode (5.02 cm) and length-to-mass shoot (97.8 cm g−1) and root (more than 140 m g−1) ratios. This may be interpreted as maximizing competition for light but also as a compensation for low shoot efficiency due to the lowest (p < 0.05) recorded values of optically-measured chlorophyll content index (22.8). Ciclope invests in biomass with highest shoot (0.06 g) and root (0.04 g) mass and a thicker root system (average diameter 0.34 mm vs. 0.29 in Saragolle) as well as a highest root/shoot ratio (0.95 g g−1 vs. 0.54 in Saragolle). Rhizosheath mass ranged between 22.14 times that of shoot mass in Ciclope and 43.40 in Saragolle (different for p < 0.05). Inoculation with Trichoderma increased the amount of rhizosheath from 9.4% in Ciclope to 36.1% in Simeto and modified root architecture in this variety more than in others. Ours are the first data on roots and seedling shoot traits of Saragolle Lucana and of Trichoderma inoculation effects on rhizosheath. This opens to new unreported interpretations of effects of Trichoderma inoculation on improving plant growth.

Screening of Soybean Genotypes Based on Root Morphology and Shoot Traits Using the Semi-Hydroponic Phenotyping Platform and Rhizobox Technique

Root-system architecture is vital for improving soybean (Glycine max L.) growth and nutrient uptake. We characterised root-system architecture and shoot traits of 30 soybean genotypes in a semi-hydroponic system 35 days after sowing (DAS) and validated eight genotypes with contrasting root-system architecture in 1.5 m-deep rhizoboxes at the flowering stage. Among them, two genotypes were selected for evaluation through to maturity. Abundant variation (coefficient of variation values ≥ 0.25) was observed in 11 of 13 measured roots and shoot traits during the early growth stage. After late growth stages, strong positive correlations were found between root traits and shoot traits, except for specific root length and diameter. Seed yield and yield traits at final harvest significantly differed between two contrasting soybean genotypes. The large-rooted genotype had a higher harvest index than the small-rooted genotype. Soybean genotypes with larger root systems had a long time to flowering than those with smaller root systems. Genotypes with large-root systems had 106% more leaf area, and 245% more shoot dry weight than those with small systems, presumably due to high canopy photosynthesis to supply the demand for carbon assimilates to roots. Total root length, and root: shoot ratio-traits data collected in the rhizobox study, strongly correlated with the same traits in the semi-hydroponic phenotyping system. We found genetic variation and phenotypic plasticity in other root and shoot traits such as taproot depth, root dry weight, specific root length, and average root diameter among the tested genotypes. Phenology, particularly time to flowering, was associated with root system size. Some root and shoot traits in the semi-hydroponic phenotyping system at the seedling stage produced similar rankings at the later phenological (flowering) stage when grown in the soil-filled rhizoboxes. The soybean genotypes characterised by vastly different root traits could be used for further glasshouse and field studies to improve adaptation to drought and other specific environments.

Rock Fragment Content in Soils Shift Root Foraging Behavior in Xerophytic Species

Aims Root traits associated with resource foraging, including fine-root branching intensity, root hair and mycorrhiza, may change in soils with various physical structure indicated by rock fragment content (RFC), while how these traits covariate at the level of individual root branching order is largely unknown.Methods We subjected two xerophytic species, Artemisia vestita (subshrub) and Bauhinia brachycarpa (shrub), to increasing RFC gradients (0%, 25%, 50% and 75%, v v-1) in an arid environment and measured fine-root traits related to resource foraging.Results Root hair density and mycorrhizal colonization of both species decreased with increasing root order, but increased in 3rd- and 4th-order roots at high RFCs (50% or 75%). The two species tend to produce more root hairs than mycorrhizas under the high RFCs. For both species, root hair density and mycorrhizal colonization intensity were negatively correlated with root length and root diameter. Rockiness reduced root branching intensity in both species comparing with rock-free soil. At the same level of RFC, A. vestita had thicker roots and lower branching intensity than B. brachycarpa, and tended to produce more root hairs.Conclusion Our results suggest the high RFC soil conditions stimulated greater foraging functions in higher root orders. We found evidence for a greater investment in root hairs and mycorrhizal symbioses as opposed to building an extensive root system in rocky soils. The subshrub and shrub species took different approaches to foraging in the rocky soil through distinctive trait syndromes of fine-root components.

Roots With Larger Specific Root Length and C: N Ratio Sustain More Complex Nematode Community

Aims Roots bridge above and belowground systems, and play a pivotal role in structuring root-associated organisms via influencing food resources and habitat conditions. Most studies focused on the relationships between plant identity and root-associated organisms, however, little is known about how root traits affect nematode communities within the rhizosphere. Methods We investigated the relationships between root traits of four plant species and nematode diversity, community structure and trophic complexity in an ex-arable field. Results While the relative abundance of herbivorous nematodes was negatively associated with specific root length (SRL), specific root area (SRA), root length density (RLD) and root C: N ratio, free-living nematodes were positively affected by these traits, implying a multifaceted effect of root traits on root-associated organisms. Importantly, we found that finer root systems promoted the complexity of the nematode community, by increasing the relative abundance of high trophic-level nematodes (i.e., omnivores and predators) and enhancing nematode diversity. Conclusion Our findings suggest that root traits could be reliable indicators of soil community structure and interactions, and provide new insights into soil biodiversity and functional maintenance.

Characterization of Root System Architecture Traits in Diverse Soybean Genotypes Using a Semi-Hydroponic System

Phenotypic variation and correlations among root traits form the basis for selecting and breeding soybean varieties with efficient access to water and nutrients and better adaptation to abiotic stresses. Therefore, it is important to develop a simple and consistent system to study root traits in soybean. In this study, we adopted the semi-hydroponic system to investigate the variability in root morphological traits of 171 soybean genotypes popularized in the Yangtze and Huaihe River regions, eastern China. Highly diverse phenotypes were observed: shoot height (18.7–86.7 cm per plant with a median of 52.3 cm); total root length (208–1663 cm per plant with a median of 885 cm); and root mass (dry weight) (19.4–251 mg per plant with a median of 124 mg). Both total root length and root mass exhibited significant positive correlation with shoot mass (p ≤ 0.05), indicating their relationship with plant growth and adaptation strategies. The nine selected traits contributed to one of the two principal components (eigenvalues > 1), accounting for 78.9% of the total genotypic variation. Agglomerative hierarchical clustering analysis separated the 171 genotypes into five major groups based on these root traits. Three selected genotypes with contrasting root systems were validated in soil-filled rhizoboxes (1.5 m deep) until maturity. Consistent ranking of the genotypes in some important root traits at various growth stages between the two experiments indicates the reliability of the semi-hydroponic system in phenotyping root trait variability at the early growth stage in soybean germplasms.

Root Trait Diversity in Field Grown Durum Wheat and Comparison with Seedlings

Roots are important for crop adaptation, particularly in dryland environments. We evaluated root development of 37 durum wheat genotypes (modern cultivars and landraces) in the field at the adult plant stage, through a shovelomics approach. Large genotypic variability was found for root traits. Differences between the landraces and modern cultivars were the main driver of this variation, with landraces showing higher plant vigor for roots and shoots. Nonetheless, genotypic variation within groups was also observed, related to different models of root growth, largely independent of total root length. These two models represented root growth were oriented either to occupy more soil volume, or to occupy less soil volume with increased density. The field results were then compared with root data previously collected in seedlings using a filter paper-based method, to assess whether early root anticipated adult root features. Field plants showed a narrower root angle than seedlings. In particular, landraces presented a narrower root angle than cultivars, but only at seedling stage. Potentially useful correlations were found between the two growth stages for root length and number.