Regulatory networks controlling the development of the root system and the formation of lateral roots: a comparative analysis of the roles of pericycle and vascular cambium.

Background: The production of a new lateral root from parental root primary tissues has been investigated extensively, and the most important regulatory mechanisms are now well known. A first regulatory mechanism is based on the synthesis of small peptides which interact ectopically with membrane receptors to elicit a modulation of transcription factor target genes. A second mechanism involves a complex cross-talk between plant hormones. It is known that lateral roots are formed even in parental root portions characterized by the presence of secondary tissues, but there is not yet agreement about the putative tissue source providing the cells competent to become founder cells of a new root primordium. Scope: We suggest models of possible regulatory mechanisms for inducing specific root vascular cambium (VC) stem cells to abandon their activity in the production of xylem and phloem elements and to start instead the construction of a new lateral root primordium. Considering the ontogenic nature of the VC, the models which we suggest are the result of a comparative review of mechanisms known to control the activity of stem cells in the root apical meristem, procambium and VC. Stem cells in the root meristems can inherit various competences to play different roles, and their fate could be decided in response to cross-talk between endogenous and exogenous signals. Conclusions: We have found a high degree of relatedness among the regulatory mechanisms controlling the various root meristems. This fact suggests that competence to form new lateral roots can be inherited by some stem cells of the VC lineage. This kind of competence could be represented by a sensitivity of specific stem cells to factors such as those presented in our models.

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.

Acclimation of fine root respiration to soil warming involves starch deposition in very fine and fine roots: a case study in Fagus sylvatica saplings.

Root activities in terms of respiration and non-structural carbohydrates (NSC) storage and mobilization have been suggested as major physiological roles in fine root lifespan. As more frequent heat waves and drought periods within the next decades are expected, to what extent does thermal acclimation in fine roots represent a mechanism to cope with such upcoming climatic conditions? In this study, the possible changes in very fine (diameter < 0.5 mm) and fine (0.5-1 mm) root morphology and physiology in terms of respiration rate and NSC [soluble sugars (SS) and starch] concentrations, were investigated on 2-year-old Fagus sylvatica saplings subjected to a simulated long-lasting heat wave event and to co-occurring soil drying. For both very fine and fine roots, soil temperature (ST) resulted inversely correlated with specific root length, respiration rates and SSs concentration, but directly correlated with root mass, root tissue density and starch concentration. In particular, starch concentration increased under 28 °C for successively decreasing under 21 °C ST. These findings showed that thermal acclimation in very fine and fine roots due to 24 days exposure to high ST (∼ 28 °C), induced starch accumulation. Such 'carbon-savings strategy' should bear the maintenance costs associated to the recovery process in case of restored favorable environmental conditions, such as those occurring at the end of a heat wave event. Drought condition seems to affect the fine root vitality much more under moderate than high temperature condition, making the temporary exposure to high ST less threatening to root vitality than expected.

MicroRNAs expression patterns in the response of poplar woody root to bending stress.

MAIN CONCLUSION: The paper reports for the first time, in poplar woody root, the expression of five mechanically-responsive miRNAs. The observed highly complex expression pattern of these miRNAs in the bent root suggest that their expression is not only regulated by tension and compression forces highlighting their role in several important processes, i.e., lateral root formation, lignin deposition, and response to bending stress. Mechanical stress is one of the major abiotic stresses significantly affecting plant stability, growth, survival, and reproduction. Plants have developed complex machineries to detect mechanical perturbations and to improve their anchorage. MicroRNAs (miRNAs), small non-coding RNAs (18-24 nucleotides long), have been shown to regulate various stress-responsive genes, proteins and transcription factors, and play a crucial role in counteracting adverse conditions. Several mechanical stress-responsive miRNAs have been identified in the stem of Populus trichocarpa plants subjected to bending stress. However, despite the pivotal role of woody roots in plant anchorage, molecular mechanisms regulating poplar woody root responses to mechanical stress have still been little investigated. In the present paper, we investigate the spatial and temporal expression pattern of five mechanically-responsive miRNAs in three regions of bent poplar woody taproot and unstressed controls by quantitative RT-PCR analysis. Alignment of the cloned and sequenced amplified fragments confirmed that their nucleotide sequences are homologous to the mechanically-responsive miRNAs identified in bent poplar stem. Computational analysis identified putative target genes for each miRNA in the poplar genome. Additional miRNA target sites were found in several mechanical stress-related factors previously identified in poplar root and a subset of these was further analyzed for expression at the mRNA or protein level. Integrating the results of miRNAs expression patterns and target gene functions with our previous morphological and proteomic data, we concluded that the five miRNAs play crucial regulatory roles in reaction woody formation and lateral root development in mechanically-stressed poplar taproot.

Temporal analysis of poplar woody root response to bending stress.

Temperate perennial woody plants use different environmental signals to coordinate their growth and development in relation to seasonal changes. Preliminary evidences suggest that, even during dormancy, plants maintain effective metabolic activities and molecular mechanisms ensuring them an eventual recording of mechanical loads during winter times. Despite their great importance for productivity and survival, plant biology investigations have poorly characterized the root growth cycle and its response to environmental stresses. In this study, we describe the proteomic changes occurring over the time in poplar root either in the absence or in response to a bending stress; corresponding expression of cell cycle regulator and auxin transporter genes was also evaluated by reverse transcription polymerase chain reaction analysis. Our results confirm previous evidences on the effect of the bending stress on the anticipation of root growth resumption, providing additional insights on a temporal modulation of various plant metabolic processes involved in dormancy break, growth resumption and stress response in the bent root; these events seem related to the differential compression and tension force distribution occurring over the plant taproot.

The adaptability of Ulmus pumila and the sensitivity of Populus sibirica to semi-arid steppe is reflected in the stem and root vascular cambium and anatomical wood traits.

Afforestation success is measured by the tree establishment and growth capacity which contribute to a range of ecosystem services. In the Mongolian steppe, Populus sibirica and Ulmus pumila have been tested as candidate species for large afforestation programs, by analyzing their response to a combination of irrigation and fertilization treatments. While in temperate and Mediterranean forest ecosystems, xylogenetic studies provide insight into the trees' plasticity and adaptability, this type of knowledge is non-existent in semi-arid regions, whose climatic features are expected to become a global issue. Furthermore, in general, a comparison between the stem and root response is scarce or absent. In the present study, we show that the anatomical traits of the vascular cambium and the xylem, from stem and root microcores, reflect the previously noted dependence of P. sibirica from irrigation - as they proportionally increase and the higher adaptability of U. pumila to drought - due to the reduced impact across all five characteristics. As the first wood anatomy study of these species in semiarid areas, future research is urgently needed, as it could be a tool for quicker understanding of species' suitability under expected to be exacerbated semi-arid conditions.

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.

Plant Growth in LED-Sourced Biophilic Environments Is Improved by the Biochar Amendment of Low-Fertility Soil, the Reflection of Low-Intensity Light, and a Continuous Photoperiod.

Introducing plants in the design of biophilic indoor environments is fundamental for improving human health, well-being, and performance. Previous studies showed that the phenotype of the model plant Arabidopsis thaliana grown under LED-sourced CoeLux® lighting systems was characterized by low biomass production rates, a small leaf area, and a low lamina-to-petiole length ratio, suggesting the onset of a strong shade avoidance syndrome. Therefore, it is essential to identify new strategies to improve plant growth under these peculiar light conditions. In the present work, we investigated the effects of two growing media (i.e., low-fertility soil and soil-less substrate), solid and liquid fertilizers, manure, biochar, perlite, mirror reflection of light, and a 24 h photoperiod on A. thaliana plants growing under CoeLux® lighting systems at a light intensity of 30 µmol m-2s-1. We found that the biochar soil amendment to low-fertility soil increases both the above-ground plant biomass and leaf area. Furthermore, the application of a mirror behind the plants and a continuous photoperiod improves not only the biomass and the leaf area but also the lamina-to-petiole length ratio. The combination of different beneficial treatments can further boost plant growth in the low-intensity light environment characterizing the CoeLux® biophilic lighting systems.

Light spectra of biophilic LED-sourced system modify essential oils composition and plant morphology of Mentha piperita L. and Ocimum basilicum L.

Investigating morphological and molecular mechanisms that plants adopt in response to artificial biophilic lighting is crucial for implementing biophilic approaches in indoor environments. Also, studying the essential oils (EOs) composition in aromatic plants can help unveil the light influence on plant metabolism and open new investigative routes devoted to producing valuable molecules for human health and commercial applications. We assessed the growth performance and the EOs composition of Mentha x piperita and Ocimum basilicum grown under an innovative artificial biophilic lighting system (CoeLux®), that enables the simulation of natural sunlight with a realistic sun perception, and compared it to high-pressure sodium lamps (control) We found that plants grown under the CoeLux® light type experienced a general suppression of both above and belowground biomass, a high leaf area, and a lower leaf thickness, which might be related to the shade avoidance syndrome. The secondary metabolites composition in the plants' essential oils was scarcely affected by both light intensity and spectral composition of the CoeLux® light type, as similarities above 80% were observed with respect to the control light treatments and within both plant species. The major differences were detected with respect to the EOs extracted from plants grown under natural sunlight (52% similarity in M. piperita and 75% in O. basilicum). Overall, it can be speculated that the growth of these two aromatic plants under the CoeLux® lighting systems is a feasible strategy to improve biophilic approaches in closed environments that include both plants and artificial sunlight. Among the two plant species analyzed, O. basilicum showed an overall better performance in terms of both morphological traits and essential oil composition. To increase biomass production and enhance the EOs quality (e.g., higher menthol concentrations), further studies should focus on technical solutions to raise the light intensity irradiating plants during their growth under the CoeLux® lighting systems.

The proteome of Populus nigra woody root: response to bending.

BACKGROUND AND AIMS: Morphological and biomechanical alterations occurring in woody roots of many plant species in response to mechanical stresses are well documented; however, little is known about the molecular mechanisms regulating these important alterations. The first forest tree genome to be decoded is that of Populus, thereby providing a tool with which to investigate the mechanisms controlling adaptation of woody roots to changing environments. The aim of this study was to use a proteomic approach to investigate the response of Populus nigra woody taproot to mechanical stress. METHODS: To simulate mechanical perturbations, the taproots of 30 one-year-old seedlings were bent to an angle of 90 ° using a steel net. A spatial and temporal two-dimensional proteome map of the taproot axis was obtained. We compared the events occurring in the above-bending, central bending and below-bending sectors of the taproot. KEY RESULTS: The first poplar woody taproot proteome map is reported here; a total of 207 proteins were identified. Spatial and temporal proteomic analysis revealed that factors involved in plant defence, metabolism, reaction wood formation and lateral root development were differentially expressed in the various sectors of bent vs. control roots, seemingly in relation to the distribution of mechanical forces along the stressed woody taproots. A complex interplay among different signal transduction pathways involving reactive oxygen species appears to modulate these responses. CONCLUSIONS: Poplar woody root uses different temporal and spatial mechanisms to respond to mechanical stress. Long-term bending treatment seem to reinforce the defence machinery, thereby enabling the taproot to better overcome winter and to be ready to resume growth earlier than controls.

Involvement of lignin and hormones in the response of woody poplar taproots to mechanical stress.

Mechanical stress is a widespread condition caused by numerous environmental factors that severely affect plant stability. In response to mechanical stress, plants have evolved complex response pathways able to detect mechanical perturbations and inducing a suite of modifications in order to improve anchorage. The response of woody roots to mechanical stresses has been studied mainly at the morphological and biomechanical level, whereas investigations on the factors triggering these important alterations are still at the initial stage. Populus has been widely used to study the response of stem to different mechanical stresses and, since it has the first forest tree genome to be decoded, represents a model woody plant for addressing questions on the mechanisms controlling adaptation of woody roots to changing environments. In this study, a morphological and physiological analysis was used to investigate factors controlling modifications in Populus nigra woody taproots subjected to mechanical stress. An experimental model analyzing spatial and temporal mechanical force distribution along the woody taproot axis enabled us to compare the events occurring in its above-, central- and below-bending sectors. Different morphogenetic responses and local variations of lignin and plant hormones content have been observed, and a relation with the distribution of the mechanical forces along the stressed woody taproots is hypothesized. We investigated the differences of the response to mechanical stress induction during the time; in this regard, we present data referring to the effect of mechanical stress on plant transition from its condition of winter dormancy to that of full vegetative activity.

Photoreceptors' gene expression of Arabidopsis thaliana grown with biophilic LED-sourced lighting systems.

Using specific photoreceptors, plants can sense light signals fundamental to their growth and development under changing light conditions. Phytochromes sense red and far-red light, cryptochromes and phototropins sense UV-A and blue light, while the UVR8 gene senses UV-B signals. The study of the molecular mechanisms used by plants to respond to artificial biophilic lighting is of pivotal importance for the implementation of biophilic approaches in indoor environments. CoeLux® is a new lighting system that reproduces the 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. We used the model plant Arabidopsis thaliana to assess the gene expression of the main plant photoreceptors at different light intensities and at different times after exposure to the CoeLux® light type, using high-pressure sodium (HPS) lamps as control light type. Genes belonging to different families of photoreceptors showed a similar expression pattern, suggesting the existence of a common upstream regulation of mRNA transcription. In particular, PHYA, PHYC, PHYD, CRY1, CRY2, PHOT1, and UVR8, showed a common expression pattern with marked differences between the two light types applied; under the HPS light type, the expression levels are raising with the decrease of light intensity, while under the CoeLux® light type, the expression levels remain nearly constant at a high fold. Moreover, we showed that under biophilic illumination the light spectrum plays a crucial role in the response of plants to light intensity, both at the molecular and morphological levels.

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.

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.

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.

Meristematic Connectome: A Cellular Coordinator of Plant Responses to Environmental Signals?

Mechanical stress in tree roots induces the production of reaction wood (RW) and the formation of new branch roots, both functioning to avoid anchorage failure and limb damage. The vascular cambium (VC) is the factor responsible for the onset of these responses as shown by their occurrence when all primary tissues and the root tips are removed. The data presented confirm that the VC is able to evaluate both the direction and magnitude of the mechanical forces experienced before coordinating the most fitting responses along the root axis whenever and wherever these are necessary. The coordination of these responses requires intense crosstalk between meristematic cells of the VC which may be very distant from the place where the mechanical stress is first detected. Signaling could be facilitated through plasmodesmata between meristematic cells. The mechanism of RW production also seems to be well conserved in the stem and this fact suggests that the VC could behave as a single structure spread along the plant body axis as a means to control the relationship between the plant and its environment. The observation that there are numerous morphological and functional similarities between different meristems and that some important regulatory mechanisms of meristem activity, such as homeostasis, are common to several meristems, supports the hypothesis that not only the VC but all apical, primary and secondary meristems present in the plant body behave as a single interconnected structure. We propose to name this structure "meristematic connectome" given the possibility that the sequence of meristems from root apex to shoot apex could represent a pluricellular network that facilitates long-distance signaling in the plant body. The possibility that the "meristematic connectome" could act as a single structure active in adjusting the plant body to its surrounding environment throughout the life of a plant is now proposed.

Effects of Irrigation and Fertilization on the Morphophysiological Traits of Populus sibirica Hort. Ex Tausch and Ulmus pumila L. in the Semiarid Steppe Region of Mongolia.

Desertification is impeding the implementation of reforestation efforts in Mongolia. Many of these efforts have been unsuccessful due to a lack of technical knowledge on water and nutrient management strategies, limited financial support, and short-lived rainfall events. We investigated the effects of irrigation and fertilization on the morphophysiological traits of Populus sibirica Hort. Ex Tausch and Ulmus pumila L. and to suggest irrigation and fertilization strategies for reforestation. Different irrigation and fertilizer treatments were applied: no irrigation, 2 L h-1, 4 L h-1, and 8 L h-1 of water; no fertilizer, 2 L h-1 + NPK, 4 L h-1 + NPK, and 8 L h-1 + NPK; and no compost, 2 L h-1 + compost, 4 L h-1 + compost, and 8 L h-1 + compost. The leaf area (LA) and specific leaf area (SLA) of both species responded positively to 4 and 8 L h-1. Results also showed that the addition of either NPK or compost to 4 or 8 L h-1 irrigation resulted in a higher LA, SLA, and leaf biomass (LB). Total chlorophyll content decreased with irrigation in both species. The same pattern was detected when a higher amount of irrigation was combined with fertilizers. Lastly, we found that both diurnal and seasonal leaf water potential of plants grown in 4 or 8 L h-1 were significantly higher than those of plants grown in control plots. Therefore, 4 or 8 L h-1 with either NPK or compost has shown to be the optimal irrigation and fertilization strategy for the species in an arid and semiarid region of Mongolia. Results should provide us with a better understanding of tree responses to varying amounts of irrigation with or without fertilizer in pursuit of sustainable forest management in arid and semiarid ecosystems.

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.

The proteome of lentil (Lens culinaris Medik.) seeds: discriminating between landraces.

Lentil (Lens culinaris Medik.) is one of the most ancient crops of the Mediterranean region used for human nutrition; an extensive differentiation of L. culinaris over millennia has resulted in a number of different landraces. As a consequence of environmental and socio-economic issues, the disappearance of many of them occurred in more recent times. To investigate the potential of proteomics as a tool in phylogenetic studies, testing the possibility to identify specific markers of different plant landraces, 2-D gel electrophoretic maps of mature seeds were obtained from seven lentil populations belonging to a local ecotype (Capracotta) and five commercial varieties (Turca Rossa, Canadese, Castelluccio di Norcia, Rascino and Colfiorito). 2-DE analysis resolved hundreds of protein species in each lentil sample, among which only 122 were further identified by MALDI-TOF PMF and/or nanoLC-ESI-LIT-MS/MS, probably as a result of the poor information available on L. culinaris genome. A comparison of these maps revealed that 103 protein spots were differentially expressed within and between populations. The multivariate statistical analyses carried out on these variably expressed spots showed that 24 protein species were essential for population discrimination, thus determining their proposition as landrace markers. Besides providing the first reference map of mature lentil seeds, our data confirm previous studies based on morphological/genetic observations and further support the valuable use of proteomic techniques as phylogenetic tool in plant studies.