Terrestrial laser scanning and low magnetic field digitization yield similar architectural coarse root traits for 32-year-old Pinus ponderosa trees.

BACKGROUND: Understanding how trees develop their root systems is crucial for the comprehension of how wildland and urban forest ecosystems plastically respond to disturbances such as harvest, fire, and climate change. The interplay between the endogenously determined root traits and the response to environmental stimuli results in tree adaptations to biotic and abiotic factors, influencing stability, carbon allocation, and nutrient uptake. Combining the three-dimensional structure of the root system, with root morphological trait information promotes a robust understanding of root function and adaptation plasticity. Low Magnetic Field Digitization coupled with AMAPmod (botAnique et Modelisation de l'Architecture des Plantes) software has been the best-performing method for describing root system architecture and providing reliable measurements of coarse root traits, but the pace and scale of data collection remain difficult. Instrumentation and applications related to Terrestrial Laser Scanning (TLS) have advanced appreciably, and when coupled with Quantitative Structure Models (QSM), have shown some potential toward robust measurements of tree root systems. Here we compare, we believe for the first time, these two methodologies by analyzing the root system of 32-year-old Pinus ponderosa trees. RESULTS: In general, at the total root system level and by root-order class, both methods yielded comparable values for the root traits volume, length, and number. QSM for each root trait was highly sensitive to the root size (i.e., input parameter PatchDiam) and models were optimized when discrete PatchDiam ranges were specified for each trait. When examining roots in the four cardinal direction sectors, we observed differences between methodologies for length and number depending on root order but not volume. CONCLUSIONS: We believe that TLS and QSM could facilitate rapid data collection, perhaps in situ, while providing quantitative accuracy, especially at the total root system level. If more detailed measures of root system architecture are desired, a TLS method would benefit from additional scans at differing perspectives, avoiding gravitational displacement to the extent possible, while subsampling roots by hand to calibrate and validate QSM models. Despite some unresolved logistical challenges, our results suggest that future use of TLS may hold promise for quantifying tree root system architecture in a rapid, replicable manner.

An Integrated Multilevel Approach Unveils Complex Seed-Nanoparticle Interactions and Their Implications for Seed Priming.

Nanotechnology has the potential to revolutionize agriculture with the introduction of engineered nanomaterials. However, their use is hindered by high cost, marginal knowledge of their interactions with plants, and unpredictable effects related to massive use in crop cultivation. Nanopriming is an innovative seed priming technology able to match economic, agronomic, and environmental needs in agriculture. The present study was focused on unveiling, by a multilevel integrated approach, undisclosed aspects of seed priming mediated by iron oxide magnetic nanoparticles in pepper seeds (Capsicum annuum), one of the most economically important crops worldwide. Inductively coupled plasma atomic emission mass spectrometry and scanning electron microscopy were used to quantify the MNP uptake and assess seed surface changes. Magnetic resonance imaging mapped the distribution of MNPs prevalently in the seed coat. The application of MNPs significantly enhanced the root and vegetative growth of pepper plants, whereas seed priming with equivalent Fe concentrations supplied as FeCl3 did not yield these positive effects. Finally, global gene expression by RNA-sequencing identified more than 2,200 differentially expressed genes, most of them involved in plant developmental processes and defense mechanisms. Collectively, these data provide evidence on the link between structural seed changes and an extensive transcriptional reprogramming, which boosts the plant growth and primes the embryo to cope with environmental challenges that might occur during the subsequent developmental and growth stages.

Counteracting action of Bacillus stratosphericus and Staphylococcus succinus strains against deleterious salt effects on Zea mays L.

The salinization of soil is the process of progressive accumulation of salts such as sulfates, sodium, or chlorides into the soil. The increased level of salt has significant effects on glycophyte plants, such as rice, maize, and wheat, which are staple foods for the world's population. Consequently, it is important to develop biotechnologies that improve crops and clean up the soil. Among other remediation methods, there is an environmentally friendly approach to ameliorate the cultivation of glycophyte plants in saline soil, namely, the use of microorganisms tolerant to salt with growth-promoting features. Plant growth-promoting rhizobacteria (PGPR) can improve plant growth by colonizing their roots and playing a vital role in helping plants to establish and grow in nutrient-deficient conditions. Our research aimed to test in vivo halotolerant PGPR, isolated and characterized in vitro in a previous study conducted in our laboratory, inoculating them on maize seedlings to improve their growth in the presence of sodium chloride. The bacterial inoculation was performed using the seed-coating method, and the produced effects were evaluated by morphometric analysis, quantization of ion contents (sodium, potassium), produced biomass, both for epigeal (shoot) and hypogeal (root) organs, and by measuring salt-induced oxidative damage. The results showed an increase in biomass and sodium tolerance and even a reduction of oxidative stress in seedlings pretreated with a PGPR bacterial consortium (Staphylococcus succinus + Bacillus stratosphericus) over the control. Moreover, we observed that salt reduces growth and alters root system traits of maize seedlings, while bacterial treatment improves plant growth and partially restores the root architecture system in saline stress conditions. Therefore, the PGPR seed-coating or seedling treatment could be an effective strategy to enhance sustainable agriculture in saline soils due to the protection of the plants from their inhibitory effect.

Unusual late-fall wildfire in a pre-Alpine Fagus sylvatica forest reduced fine roots in the shallower soil layer and shifted very fine-root growth to deeper soil depth.

After an unusual, late-fall wildfire in a European beech (Fagus sylvatica L.) forest in the pre-Alps of northern Italy, the finest roots (0‒0.3 mm diameter) were generally the most responsive to fire, with the effect more pronounced at the shallowest soil depth. While roots 0.3‒1 mm in diameter had their length and biomass at the shallowest soil depth reduced by fire, fire stimulated more length and biomass at the deepest soil depth compared to the control. Fire elevated the total length of dead roots and their biomass immediately and this result persisted through the first spring, after which control and fire-impacted trees had similar fine root turnover. Our results unveiled the fine-root response to fire when subdivided by diameter size and soil depth, adding to the paucity of data concerning fire impacts on beech roots in a natural condition and providing the basis for understanding unusual fire occurrence on root traits. This study suggests that F. sylvatica trees can adapt to wildfire by plastically changing the distribution of fine-root growth, indicating a resilience mechanism to disturbance.

Fertilization reduces root architecture plasticity in Ulmus pumila used for afforesting Mongolian semi-arid steppe.

In this study, we assessed the functional and architectural traits in the coarse roots of Ulmus pumila trees, which are used for afforesting the semi-arid steppe of Mongolia. Tree growth was supported by different watering regimes (no watering, 2, 4, and 8 L h-1) and by two types of soil fertilization (NPK and compost). In July, 2019, for each of these treatments six trees, outplanted in 2011 as 2-year-old seedlings from a container nursery, were randomly selected, excavated by hand, and digitized. The build-up of root length correlated positively with increasing levels of watering for both soil depths analyzed. The application of fertilizers led to root growth suppression resulting in a general reduction of root length in a lowered rooting depth. When root system characteristics were analyzed in relation to wind direction, unfertilized trees showed higher root diameter values in both soil layers of leeward quadrants, likely a response to mechanical forces to improve stability. On the contrary, fertilized trees did not show differences in root diameter among the different quadrants underscoring a strong reduction in root plasticity with a lack of morpho-architectural response to the mechanical forces generated by the two prevailing winds. Finally, the root branching density, another important trait for fast dissipation of mechanical forces, was significantly reduced by the fertilization, independently of the quadrants and watering regime. Our results suggest that knowledge of the root response to the afforestation techniques applied in the semi-arid steppe of Mongolia is a necessary step for revealing the susceptibility of this forest shelterbelt to the exacerbating environmental conditions caused by climate change and, thus, to the development of a sustainable and successful strategy to restore degraded lands.

The perils of planning strategies to increase vitamin C content in plants: Beyond the hype.

Ever since the identification of vitamin C (ascorbic acid, AsA) as an essential molecule that humans cannot synthesize on their own, finding adequate dietary sources of AsA became a priority in nutrition research. Plants are the main producers of AsA for humans and other non-synthesizing animals. It was immediately clear that some plant species have more AsA than others. Further studies evidenced that AsA content varies in different plant organs, in different developmental stages/environmental conditions and even within different cell compartments. With the progressive discovery of the genes of the main (Smirnoff-Wheeler) and alternative pathways coding for the enzymes involved in AsA biosynthesis in plants, the simple overexpression of those genes appeared a suitable strategy for boosting AsA content in any plant species or organ. Unfortunately, overexpression experiments mostly resulted in limited, if any, AsA increase, apparently due to a tight regulation of the biosynthetic machinery. Attempts to identify regulatory steps in the pathways that could be manipulated to obtain unlimited AsA production were also less successful than expected, confirming the difficulties in "unleashing" AsA synthesis. A different approach to increase AsA content has been the overexpression of genes coding for enzymes catalyzing the recycling of the oxidized forms of vitamin C, namely monodehydroascorbate and dehydroascorbate reductases. Such approach proved mostly effective in making the overexpressors apparently more resistant to some forms of environmental stress, but once more did not solve the issue of producing massive AsA amounts for human diet. However, it should also be considered that a hypothetical unlimited increase in AsA content is likely to interfere with plant development, which is in many ways regulated by AsA availability itself. The present review article aims at summarizing the many attempts made so far to improve AsA production/content in plants, evidencing the most promising ones, and at providing information about the possible unexpected consequences of a pure biotechnological approach not keeping into account the peculiar features of the AsA system in plants.

Morpho-Physiological Responses of Arabidopsis thaliana L. to the LED-Sourced CoeLux® System.

The CoeLux® lighting system reproduces the true effect of natural sunlight entering through an opening in the ceiling, with a realistic sun perceived at an infinite distance surrounded by a clear blue sky. It has already been demonstrated that this new lighting system generates long-term positive effects on human beings; however, there are no investigations so far concerning the plant responses to CoeLux® lighting. To fill this gap, the model plant Arabidopsis thaliana L. was grown at four different distances from the light source, corresponding to four different light intensities (120, 70, 30, 20 µmol m-2 s-1). High-pressure sodium lamps were used as control light. Plant phenology and morpho-physiological traits were monitored to assess for the first time the ability of plants to grow and develop under the light spectrum and intensity of the CoeLux® system. Plants grown at the lower light intensities showed a delayed life cycle and were significantly smaller than plants grown with more light. Furthermore, plants grown under the CoeLux® light type showed an additional deficit when compared to control plants. Overall, our results show that both the light spectrum and intensity of the CoeLux® system had a strong impact on A. thaliana growth performance.

Root Biomass Distribution of Populus sibirica and Ulmus pumila Afforestation Stands Is Affected by Watering Regimes and Fertilization in the Mongolian Semi-arid Steppe.

Desertification of the semi-arid steppe of Mongolia is advancing very rapidly, motivating afforestation efforts. The "Green Belt" joint project (Government of Mongolia and Republic of Korea), which aims to mitigate soil degradation and develop agroforestry activities through the planting of a forest shelterbelt, is one such response. In these plantations, tree growth has been supported by different watering regimes (no watering, 2, 4, and 8 L h-1) and by two types of soil fertilization (NPK and Compost). The present paper analyses the effect of these techniques on soil chemistry and root biomass partitioning of Populus sibirica (Horth ex Tausch) and Ulmus pumila (L.) tree species. In July 2019, at the plantation site in Lun Soum, Tuv province (Mongolia), six trees were excavated by hand in each treatment, the root system was divided into taproot and five diameter classes (0-2; 2-5; 5-10; 10-20; > 20 mm), and the biomass was measured. Soil organic matter, macronutrients, and pH were also measured. The addition of fertilizers in the long-term did not enhance the soil chemical properties. The build-up of root biomass in both species correlated positively with increasing levels of the watering, while the application of fertilizers led to root growth suppression. For most of the root classes and both species, an irrigation level of 4 L h-1 was sufficient to yield the highest biomass and could be recommended for afforesting the semi-arid steppe of Mongolia. The root biomass of P. sibirica was more dependent on the watering regimes and of U. pumila was more negatively influenced by the application of fertilizers, indicating that U. pumila, due to the its lower water need, could be suitable for afforesting semi-arid environments. Our experiments suggest that afforestation practices in the semi-arid steppe of Mongolia should be supported by a prior analysis of plants' needs, soil type, dose, and type of fertilizers to be applied. Knowledge of the root response to the supporting techniques is necessary for choosing the best one for the plantation and, thus, to develop a sustainable and successful strategy to restore these degraded lands.

Pioneer and fibrous root seasonal dynamics of Vitis vinifera L. are affected by biochar application to a low fertility soil: A rhizobox approach.

The present work analyzes the impact of biochar-induced modification of soil physico-chemical properties on intra-annual growth dynamics of pioneer and fibrous grapevine roots. A scanner inserted into a buried rhizobox with a transparent side facing the plant root system was used to acquire images of pioneer and fibrous roots of control and biochar-treated plants throughout the vegetative season. Images were analyzed with ImageJ software to measure root traits. Biochar treatment increased soil pH, nutrient concentration, and water content during the driest and warmest period, while bulk density was reduced. Analysis of both pioneer and fibrous root traits highlighted a single peak of growth during the vegetative season. Pioneer roots were thicker and grew faster than fibrous roots, which were longer and more numerous. Amelioration of physico-chemical properties of biochar-amended soil stimulated an earlier root lengthening, and a higher root number at the onset of the season, which resulted in a greater canopy development compared to control plants. Later, in summer, as a consequence of the higher water content of biochar-treated soil, plants modified their root architecture, lowering the number of fibrous roots probably because of the reduced need to exploit soil for water and nutrient uptake.

Genetic Diversity and Differentiation of Eleven Medicago Species from Campania Region Revealed by Nuclear and Chloroplast Microsatellites Markers.

The species belonging to the genus Medicago are considered a very important genetic resource at global level both for planet's food security and for sustainable rangelands management. The checklist of the Italian flora (2021) includes a total number of 40 Medicago species for Italy, and 27 for Campania region, with a number of doubtful records or related to species no more found in the wild. In this study, 10 Medicago species native to Campania region, and one archaeophyte (M. sativa), identified by means of morphological diagnostic characters, were analyzed in a blind test to assay the efficacy of nine microsatellite markers (five cp-SSRs and four n-SSRs). A total number of 33 individuals from 6 locations were sampled and genotyped. All markers were polymorphic, 40 alleles were obtained with n-SSRs ranging from 8-12 alleles per locus with an average of 10 alleles per marker, PIC values ranged from 0.672 to 0.847, and the most polymorphic SSR was MTIC 564. The cp-SSRs markers were highly polymorphic too; PIC values ranged from 0.644 to 0.891 with an average of 0.776, the most polymorphic cp-SSR was CCMP10. 56 alleles were obtained with cp-SSRs ranging from 7 to 17 alleles per locus with an average of 11. AMOVA analysis with n-SSR markers highlighted a great level of genetic differentiation among the 11 species, with a statistically significant fixation index (FST). UPGMA clustering and Bayesian-based population structure analysis assigned these 11 species to two main clusters, but the distribution of species within clusters was not the same for the two analyses. In conclusion, our results demonstrated that the combination of the used SSRs well distinguished the 11 Medicago species. Moreover, our results demonstrated that the use of a limited number of SSRs might be considered for further genetic studies on other Medicago species.

Reaction Wood Anatomical Traits and Hormonal Profiles in Poplar Bent Stem and Root.

Reaction wood (RW) formation is an innate physiological response of woody plants to counteract mechanical constraints in nature, reinforce structure and redirect growth toward the vertical direction. Differences and/or similarities between stem and root response to mechanical constraints remain almost unknown especially in relation to phytohormones distribution and RW characteristics. Thus, Populus nigra stem and root subjected to static non-destructive mid-term bending treatment were analyzed. The distribution of tension and compression forces was firstly modeled along the main bent stem and root axis; then, anatomical features, chemical composition, and a complete auxin and cytokinin metabolite profiles of the stretched convex and compressed concave side of three different bent stem and root sectors were analyzed. The results showed that in bent stems RW was produced on the upper stretched convex side whereas in bent roots it was produced on the lower compressed concave side. Anatomical features and chemical analysis showed that bent stem RW was characterized by a low number of vessel, poor lignification, and high carbohydrate, and thus gelatinous layer in fiber cell wall. Conversely, in bent root, RW was characterized by high vessel number and area, without any significant variation in carbohydrate and lignin content. An antagonistic interaction of auxins and different cytokinin forms/conjugates seems to regulate critical aspects of RW formation/development in stem and root to facilitate upward/downward organ bending. The observed differences between the response stem and root to bending highlight how hormonal signaling is highly organ-dependent.

WOX genes expression during the formation of new lateral roots from secondary structures in Populus nigra (L.) taproot.

Despite the large amounts of data available on lateral root formation, little is known about their initiation from secondary structures. In the present work, we applied a bending treatment to Populus nigra (L.) woody taproots to induce the formation of new lateral roots. The development of lateral roots was monitored by stereomicroscopic examination of cross-sections. Tissues were sampled from the bending zone in the proximity of the vascular cambium before (time 0) and after the application of bending at three different time points (24, 48, and 72 h) and analyzed for the expression of P. nigra WOX homologs. The initiation of new lateral roots was observed to originate from the vascular cambium zone and was followed by primordium formation and root emergence. PnWOX4a, PnWOX4b, PnWOX5a, PnWOX5b, PnWOX11/12a, and PnWOX11/12b were shown to be expressed during the formation of new lateral roots at different developmental stages. The mechanical stress simulated by bending treatment was shown to activate the molecular mechanism leading to the expression of WOX genes, which are hypothesized to control SLR formation in the cambium zone of poplar taproot.

Formation of Annual Ring Eccentricity in Coarse Roots within the Root Cage of Pinus ponderosa Growing on Slopes.

: The coarse roots of Pinus ponderosa included in the cage are the ones most involved in tree stability. This study explored the variations in traits, such as volume, cross-sectional area, and radius length of cage roots, and used those data to develop a mathematical model to better understand the type of forces occurring for each shallow lateral root segment belonging to different quadrants of the three-dimensional (3D) root system architecture. The pattern and intensity of these forces were modelled along the root segment from the branching point to the cage edge. Data of root cage volume in the upper 30 cm of soil showed a higher value in the downslope and windward quadrant while, at a deeper soil depth (> 30 cm), we found higher values in both upslope and leeward quadrants. The analysis of radius length and the cross-sectional area of the shallow lateral roots revealed the presence of a considerable degree of eccentricity of the annual rings at the branching point and at the cage edge. This eccentricity is due to the formation of compression wood, and the eccentricity changes from the top portion at the branching point to the bottom portion at the cage edge, which we hypothesize may be a response to the variation in mechanical forces occurring in the various zones of the cage. This hypothesis is supported by a mathematical model that shows how the pattern and intensity of different types of mechanical forces are present within the various quadrants of the same root system from the taproot to the cage edge.

Functional Traits of Pinus ponderosa Coarse Roots in Response to Slope Conditions.

We excavated the root systems of Pinus ponderosa trees growing on a steeply sloped, volcanic ash-influenced soil in the northern Rocky Mountains of the United States to assess their functional coarse-root traits and root system architecture. Trees, outplanted as one-year-old seedlings from a container nursery, were in their 32nd growing season on the site. We found that the trees had deployed more roots, in terms of length and volume, in the downslope and windward quadrants than in their upslope and leeward quadrants, likely a response to mechanical forces toward improving stability. Moreover, we observed the development of three types of root cages (tight, enlarged, and diffused) that likely reflect micro-site characteristics. As the cage type transitioned from tight to enlarged to diffused we measured a decrease in the overall volume of the roots associated with the cage and the taproot becoming a more prominent contributor to the overall volume of the cage. Finally, we noted the development of specialty roots, namely those with I-beam and T-beam shapes in cross section, in the downslope quadrant; these types of roots are known to better counteract compression mechanical forces. These observations improve our understanding of root plasticity and tree rooting response to environmental stimuli, which is becoming an increasingly critical topic as changes in climate increase the frequency and intensity of storms.

Non-destructive Phenotypic Analysis of Early Stage Tree Seedling Growth Using an Automated Stereovision Imaging Method.

A plant phenotyping approach was applied to evaluate growth rate of containerized tree seedlings during the precultivation phase following seed germination. A simple and affordable stereo optical system was used to collect stereoscopic red-green-blue (RGB) images of seedlings at regular intervals of time. Comparative analysis of these images by means of a newly developed software enabled us to calculate (a) the increments of seedlings height and (b) the percentage greenness of seedling leaves. Comparison of these parameters with destructive biomass measurements showed that the height traits can be used to estimate seedling growth for needle-leaved plant species whereas the greenness trait can be used for broad-leaved plant species. Despite the need to adjust for plant type, growth stage and light conditions this new, cheap, rapid, and sustainable phenotyping approach can be used to study large-scale phenome variations due to genome variability and interaction with environmental factors.

Poplar woody taproot under bending stress: the asymmetric response of the convex and concave sides.

Background and Aims Progress has been made in understanding the physiological and molecular basis of root response to mechanical stress, especially in the model plant Arabidopsis thaliana, in which bending causes the initiation of lateral root primordia toward the convex side of the bent root. In the case of woody roots, it has been reported that mechanical stress induces an asymmetric distribution of lateral roots and reaction wood formation, but the mechanisms underlying these responses are largely unknown. In the present work, the hypothesis was tested that bending could determine an asymmetric response in the two sides of the main root axis as cells are stretched on the convex side and compressed on the concave side. Methods Woody taproots of 20 seedlings were bent to an angle of 90° using a steel net. Changes in the anatomy, lignin and phytohormone content and proteome expression in the two sides of the bent root were analysed; anatomical changes, including dissimilarities and similarities to those found in poplar bent woody stem, were also considered. Key Results Compression forces at the concave side of poplar root induced the formation of reaction wood which presented a high lignin content and was associated with the induction of cambium cell activity. Auxin seemed to be the main hormone triggering lignin deposition and cell wall strengthening in the concave sides. Abscisic acid appeared to function in the water stress response induced by xylem structures and/or osmotic alterations in the compression sides, whereas gibberellins may control cell elongation and gravitropisms. Conclusions Poplar root reaction wood showed characteristics different from those produced in bent stem. Besides providing biomechanical functions, a bent root ensures water uptake and transport in the deforming condition induced by tension and compression forces by two different strategies: an increase in xylem thickness in the compressed side, and lateral root formation in the tension side.

Fine-root carbon and nitrogen concentration of European beech (Fagus sylvatica L.) in Italy Prealps: possible implications of coppice conversion to high forest.

Fine-root systems represent a very sensitive plant compartment to environmental changes. Gaining further knowledge about their dynamics would improve soil carbon input understanding. This paper investigates C and N concentrations in fine roots in relation to different stand characteristics resulting from conversion of coppiced forests to high forests. In order to evaluate possible interferences due to different vegetative stages of vegetation, fine-root sampling was repeated six times in each stand during the same 2008 growing season. Fine-root sampling was conducted within three different soil depths (0-10; 10-20; and 20-30 cm). Fine-root traits were measured by means of WinRHIZO software which enable us to separate them into three different diameter classes (0-0.5, 0.5-1.0 and 1.0-2.0 mm). The data collected indicate that N concentration was higher in converted stands than in the coppiced stand whereas C concentration was higher in the coppiced stand than in converted stands. Consequently the fine-root C:N ratio was significantly higher in coppiced than in converted stands and showed an inverse relationship with fine-root turnover rate, confirming a significant change of fine-root status after the conversion of a coppice to high forest.