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.

Effect of Polyethylene Glycol-Simulated Drought Stress on Stomatal Opening in "Modern" and "Ancient" Wheat Varieties.

Climate change is leading to an increase in the intensity, duration, and frequency of severe droughts, especially in southern and southeastern Europe, thus aggravating water scarcity problems. Water deficit stress harms the growth, physiology, and yield of crops like durum wheat. Hence, studying ancient wheat varieties' stress responses could help identify genetic traits to enhance crop tolerance to environmental stresses. In this background, this study aimed to investigate the effects of PEG 6000-stimulated drought stress in the ancient wheat variety Saragolla and the modern one Svevo by analyzing various biochemical and molecular parameters that can especially condition the stomatal movement. Our data revealed that drought stress caused a significant increase in the levels of total soluble sugars, ABA, and IAA in both selected cultivars to a greater extent in the Saragolla than in the Svevo. We demonstrated that, under water deficit stress, calcium dynamics as well as the expression of ERF109, MAPK3/6, MYB60, and TaTPC1, involved in the activation of drought-related calcium-sensitive pathways, display significant differences between the two varieties. Therefore, our study provided further evidence regarding the ability of the ancient wheat variety Saragolla to better cope with drought stress compared to the modern variety Svevo.

The physiology and anatomy study in leaves of Saragolla and Svevo wheat cultivars under polyethylene glycol-simulated drought stress.

Drought stress is increasing in frequency and severity with the progression of global climate change, thereby becoming a major concern for the growth and yield of crop plants, including wheat. The current challenge is to explore different ways of developing wheat genotypes with increased tolerance to drought. Therefore, we renewed interest in 'ancient' varieties expected to be more tolerant to environmental stress than the few elite varieties nowadays cultivated. This study aimed to perform comparative analysis of the effect of drought-simulating polyethylene glycol (PEG-6000) treatment on morpho-anatomical and physiological foliar traits of two durum wheat seedlings cultivars, Saragolla and Svevo, as these can reflect the adaptability of the plant to the environment to a certain extent. Results demonstrated that drought-stressed Saragolla leaves exhibited a greater reduction of stomatal density, a minor reduction of stomatal pore width, a wider xylem vessel mean area, greater compactness of mesophyll cells, a minor loss of chlorophyll content, as well as better photosynthetic and growth performance compared to the other variety. From such behaviours, we consider the Saragolla cultivar more drought tolerant than Svevo and therefore probably very promising for cultivation in dry areas.

Transcriptomic and proteomic analysis of a compatible tomato-aphid interaction reveals a predominant salicylic acid-dependent plant response.

BACKGROUND: Aphids are among the most destructive pests in temperate climates, causing significant damage on several crops including tomato. We carried out a transcriptomic and proteomic study to get insights into the molecular mechanisms and dynamics of the tomato response to the Macrosyphum euphorbiae aphid. RESULTS: The time course analysis of aphid infestation indicated a complex, dynamic pattern of gene expression. Several biological functions were affected and genes related to the stress and defence response were the most represented. The Gene Ontology categories of the differentially expressed genes (899) and identified proteins (57) indicated that the tomato response is characterized by an increased oxidative stress accompanied by the production of proteins involved in the detoxification of oxygen radicals. Aphids elicit a defense reaction based on the cross-communication of different hormone-related signaling pathways such as those related to the salicylic acid (SA), jasmonic acid (JA), ethylene and brassinosteroids. Among them, the SA-signaling pathway and stress-responsive SA-dependent genes play a dominant role. Furthermore, tomato response is characterized by a reduced accumulation of photosynthetic proteins and a modification of the expression of various cell wall related genes. CONCLUSIONS: Our work allowed a more comprehensive understanding of the signaling events and the defense dynamics of the tomato response to aphids in a compatible interaction and, based on experimental data, a model of the tomato-aphid molecular interaction was proposed. Considering the rapid advancement of tomato genomics, this information will be important for the development of new protection strategies.

Comparative Analysis of the Response to Polyethylene Glycol-Simulated Drought Stress in Roots from Seedlings of "Modern" and "Ancient" Wheat Varieties.

Durum wheat is widely cultivated in the Mediterranean, where it is the basis for the production of high added-value food derivatives such as pasta. In the next few years, the detrimental effects of global climate change will represent a serious challenge to crop yields. For durum wheat, the threat of climate change is worsened by the fact that cultivation relies on a few genetically uniform, elite varieties, better suited to intensive cultivation than "traditional" ones but less resistant to environmental stress. Hence, the renewed interest in "ancient" traditional varieties are expected to be more tolerant to environmental stress as a source of genetic resources to be exploited for the selection of useful agronomic traits such as drought tolerance. The aim of this study was to perform a comparative analysis of the effect and response of roots from the seedlings of two durum wheat cultivars: Svevo, a widely cultivated elite variety, and Saragolla, a traditional variety appreciated for its organoleptic characteristics, to Polyethylene glycol-simulated drought stress. The effect of water stress on root growth was analyzed and related to biochemical data such as hydrogen peroxide production, electrolyte leakage, membrane lipid peroxidation, proline synthesis, as well as to molecular data such as qRT-PCR analysis of drought responsive genes and proteomic analysis of changes in the protein repertoire of roots from the two cultivars.

An artificial intelligence-integrated analysis of the effect of drought stress on root traits of "modern" and "ancient" wheat varieties.

Due to drought stress, durum wheat production in the Mediterranean basin will be severely affected in the coming years. Durum wheat cultivation relies on a few genetically uniform "modern" varieties, more productive but less tolerant to stresses, and "traditional" varieties, still representing a source of genetic biodiversity for drought tolerance. Root architecture plasticity is crucial for plant adaptation to drought stress and the relationship linking root structures to drought is complex and still largely under-explored. In this study, we examined the effect of drought stress on the roots' characteristics of the "traditional" Saragolla cultivar and the "modern" Svevo. By means of "SmartRoot" software, we demonstrated that drought stress affected primary and lateral roots as well as root hair at different extents in Saragolla and Svevo cultivars. Indeed, we observed that under drought stress Saragolla possibly revamped its root architecture, by significantly increasing the length of lateral roots, and the length/density of root hairs compared to the Svevo cultivar. Scanning Electron Microscopy analysis of root anatomical traits demonstrated that under drought stress a greater stele area and an increase of the xylem lumen size vessel occurred in Saragolla, indicating that the Saragolla variety had a more efficient adaptive response to osmotic stress than the Svevo. Furthermore, for the analysis of root structural data, Artificial Intelligence (AI) algorithms have been used: Their application allowed to predict from root structural traits modified by the osmotic stress the type of cultivar observed and to infer the relationship stress-cultivar type, thus demonstrating that root structural traits are clear and incontrovertible indicators of the higher tolerance to osmotic stress of the Saragolla cultivar. Finally, to obtain an integrated view of root morphogenesis, phytohormone levels were investigated. According to the phenotypic effects, under drought stress,a larger increase in IAA and ABA levels, as well as a more pronounced reduction in GA levels occurred in Saragolla as compared to Svevo. In conclusion, these results show that the root growth and hormonal profile of Saragolla are less affected by osmotic stress than those of Svevo, demonstrating the great potential of ancient varieties as reservoirs of genetic variability for improving crop responses to environmental stresses.

Plasma fatty acid profile in Italian Holstein-Friesian dairy cows supplemented with natural polyphenols from the olive plant Olea Europaea L.

This study evaluated the effects of supplementing with natural functional feed on the plasma fatty acid profile of lactating Italian Holstein-Friesian dairy cows. Thirty cows in mid-lactation received the natural olive extract PHENOFEED DRY (500 mg/cow/day) which mainly comprises hydroxytyrosol, tyrosol and verbascoside. The total content of polyphenols and the antioxidant power of standard feed, enriched feed and pure extract was evaluated respectively by Folin-Ciocalteu and DPPH assay, and a characterization in HPLC-UV (High-Performance Liquid Chromatography-Ultraviolet) of bioactive molecules present in the extract PHENOFEED DRY was performed. PHENOFEED DRY was provided for 60 days, and the plasma profile of fatty acids was determined by Gas Chromatography. The administration of enriched feed resulted in an increase in the ratio of Omega-6 to Omega-3 polyunsaturated fatty acids from 3:1 to 4:1 (p<0.001). This was not influenced by the calving order. The addition of polyphenols helped to keep monounsaturated (MUFA) and saturated (SFA) levels constant and results in a significant increase in polyunsaturated (PUFA) fatty acid after 15 days of administration. The Omega-6/Omega-3 ratio was in the optimal range. The findings show that inclusion of natural functional food such as plant polyphenols helps to maintain a healthy blood fatty acid profile in lactating dairy cows.

Molecular interactions between the olive and the fruit fly Bactrocera oleae.

BACKGROUND: The fruit fly Bactrocera oleae is the primary biotic stressor of cultivated olives, causing direct and indirect damages that significantly reduce both the yield and the quality of olive oil. To study the olive-B. oleae interaction, we conducted transcriptomic and proteomic investigations of the molecular response of the drupe. The identifications of genes and proteins involved in the fruit response were performed using a Suppression Subtractive Hybridisation technique and a combined bi-dimensional electrophoresis/nanoLC-ESI-LIT-MS/MS approach, respectively. RESULTS: We identified 196 ESTs and 26 protein spots as differentially expressed in olives with larval feeding tunnels. A bioinformatic analysis of the identified non-redundant EST and protein collection indicated that different molecular processes were affected, such as stress response, phytohormone signalling, transcriptional control and primary metabolism, and that a considerable proportion of the ESTs could not be classified. The altered expression of 20 transcripts was also analysed by real-time PCR, and the most striking differences were further confirmed in the fruit of a different olive variety. We also cloned the full-length coding sequences of two genes, Oe-chitinase I and Oe-PR27, and showed that these are wound-inducible genes and activated by B. oleae punctures. CONCLUSIONS: This study represents the first report that reveals the molecular players and signalling pathways involved in the interaction between the olive fruit and its most damaging biotic stressor. Drupe response is complex, involving genes and proteins involved in photosynthesis as well as in the production of ROS, the activation of different stress response pathways and the production of compounds involved in direct defence against phytophagous larvae. Among the latter, trypsin inhibitors should play a major role in drupe resistance reaction.

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.

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.

Biochar Administration to San Marzano Tomato Plants Cultivated Under Low-Input Farming Increases Growth, Fruit Yield, and Affects Gene Expression.

Biochar is a rich-carbon charcoal obtained by pyrolysis of biomasses, which was used since antiquity as soil amendant. Its storage in soils was demonstrated contributing to abate the effects of climate changes by sequestering carbon, also providing bioenergy, and improving soil characteristics and crop yields. Despite interest in this amendant, there is still poor information on its effects on soil fertility and plant growth. Considerable variation in the plant response has been reported, depending on biomass source, pyrolysis conditions, crop species, and cultivation practices. Due to these conflicting evidences, this work was aimed at studying the effects of biochar from pyrolyzed wood at 550°C, containing 81.1% carbon and 0.91% nitrogen, on growth and yield of tomato plants experiencing low-input farming conditions. San Marzano ecotype from Southern Italy was investigated, due to its renowned quality and adaptability to sustainable farming practices. Biochar administration improved vegetative growth and berry yield, while affecting gene expression and protein repertoire in berries. Different enzymes of carbon metabolism and photosynthesis were over-represented, whereas various stress-responsive and defense proteins were down-represented. Molecular results are here discussed in relation to estimated agronomic parameters to provide a rationale justifying the growth-promoting effect of this soil amendant.

Comparative proteomic analysis of durum wheat shoots from modern and ancient cultivars.

Durum wheat is widespread cultivated in the Mediterranean basin, where it is used to produce high-quality semolina for pasta. Although over the years local and ancient wheat cultivars have been replaced by new ones, better suited to intensive cultivation, the increasing demand of consumers for nutritional and sensory qualities, as well as their attention to sustainable agronomic practices, renewed the interest toward traditional varieties. In order to fully exploit their agronomical and nutritional potential, a systematic analysis of molecular traits would be desirable. Nowadays, this examination is greatly facilitated by the current availability of high-throughput genomic and proteomic methods, which are integrated with classical measurements on plant physiology. To this purpose, we performed a comparative study on germination performances, hormone level variations, and differential protein representations of three-days germinated shoots of two traditional wheat cultivars from Southern Italy, namely Senatore Cappelli and Saragolla, and the commercial elite variety Svevo. Two-dimensional electrophoresis- and nanoLC-ESI-LIT-MS/MS-based proteomic analysis revealed 45 differentially represented spots, which were associated with 32 non-redundant protein species grouping into storage, stress/defense and metabolism/energy production functional categories. Major differences in the traditional varieties concerned over-representation of glutenins, gamma-gliadin and some enzymes of glycolysis and TCA cycle, as well as a down-representation of proteins involved in the response to stress conditions. These features were here discussed in relation to the hormone profile and the known agronomic features of traditional varieties, as compared to the commercial one.

Proteomic analysis of the major soluble components in Annurca apple flesh.

Apple is one of the most worldwide-consumed fruits and a number of cultivars, differing in organoleptic and nutritional characteristics, are available for the market. Annurca apple is a regional variety from Southern Italy, which is known for crispness, excellent taste and long shelf life of fruits. These features have renewed the interest in the investigation of their genetic potential and different studies have lead to their partial genetic and metabolic characterisation. In this study, we present the analysis of the protein repertoire of the pseudocarp tissues of three accessions of Malus x domestica Borkh. cv. Annurca, as first example of the systematic annotation of the apple proteome. Proteins were extracted from fruit tissues and resolved on 2-DE gels; commonly expressed proteins were in-gel digested and analysed by MALDI-TOF-MS and muLC-ESI-IT-MS/MS approaches. Peptide MS and MS/MS data were searched against publicly available protein and EST databases, and 44 spots were identified and associated to 28 different species. They were related to important physiological processes such as energy production, ripening and stress response. The occurrence of allergens causative of widespread food allergy syndromes was also detected. Integration of genomic, metabolomic and proteomic data will be indispensable for future molecular characterisation and hence full exploitation of the peculiar organoleptic, nutritional and agronomic traits of local cultivars of fruits.

Ophiobolin A Induces Autophagy and Activates the Mitochondrial Pathway of Apoptosis in Human Melanoma Cells.

Ophiobolin A, a fungal toxin from Bipolaris species known to affect different cellular processes in plants, has recently been shown to have anti-cancer activity in mammalian cells. In the present study, we investigated the anti-proliferative effect of Ophiobolin A on human melanoma A375 and CHL-1 cell lines. This cellular model was chosen because of the incidence of melanoma malignant tumor on human population and its resistance to chemical treatments. Ophyobolin A strongly reduced cell viability of melanoma cells by affecting mitochondrial functionality. The toxin induced depolarization of mitochondrial membrane potential, reactive oxygen species production and mitochondrial network fragmentation, leading to autophagy induction and ultimately resulting in cell death by activation of the mitochondrial pathway of apoptosis. Finally, a comparative proteomic investigation on A375 cells allowed to identify several Ophiobolin A down-regulated proteins, which are involved in fundamental processes for cell homeostasis and viability.

Effect of short-term cadmium stress on Populus nigra L. detached leaves.

Pollution by toxic metals, accumulating into soils as result of human activities, is a worldwide major concern in industrial countries. Plants exhibit different degrees of tolerance to heavy metals, as a consequence of their ability to exclude or accumulate them in particular tissues, organs or sub-cellular compartments. Molecular information about cellular processes affected by heavy metals is still largely incomplete. As a fast-growing, highly tolerant perennial plant species, poplar has become a model for environmental stress response investigations. To study the short-term effects of cadmium accumulation in leaves, we analyzed photosystem II (PSII) quantum yield, hydrogen peroxide (H2O2) generation, hormone levels variation, as well as proteome profile alteration of 50µM CdSO4 vacuum-infiltrated poplar (Populus nigra L.) detached leaves. Cadmium management brought about an early and sustained production of hydrogen peroxide, an increase of abscisic acid, ethylene and gibberellins content, as well as a decrease in cytokinins and auxin levels, whereas photosynthetic electron transport was unaffected. Proteomic analysis revealed that twenty-one proteins were differentially induced in cadmium-treated leaves. Identification of fifteen polypeptides allowed to ascertain that most of them were involved in stress response while the remaining ones were involved in photosynthetic carbon metabolism and energy production.

Tomato susceptibility to Fusarium crown and root rot: effect of grafting combination and proteomic analysis of tolerance expression in the rootstock.

Grafting can enhance the tolerance of vegetable crops to soilborne diseases. The aim of this study was to investigate whether different tomato scion-rootstock combinations may affect the plant susceptibility to Fusarium oxysporum f. sp. radicis-lycopersici (FORL), the causal agent of crown and root rot. A proteomic approach was used to investigate whenever the protein repertoire of the rootstock can be modified by FORL infection, in relation to cultivar susceptibility/tolerance to the disease. To this purpose, plants of tomato hybrids with different vigor, "Costoluto Genovese" (less vigorous) and "Kadima" (more vigorous), were grafted onto "Cuore di Bue" and "Natalia" hybrids, sensitive and tolerant versus FORL infections, respectively. Disease symptoms, plant biomasses, and protein expression patterns were evaluated 45 days after FORL inoculation. The extent of vascular discoloration caused by FORL in tomato plants grafted on "Natalia" rootstock (0.12-0.37 cm) was significantly lower than that of plants grafted on sensitive "Cuore di Bue" (1.75-6.50 cm). FORL symptoms significantly differed between "Costoluto Genovese" and "Kadima" scions only when grafted on sensitive rootstock. Shoot FW of non-inoculated "Kadima"/"Cuore di Bue" combination was 35% lower than "Kadima"/"Natalia", whereas no difference was manifested in inoculated plants. Shoot FW of inoculated "Costoluto Genovese"/"Cuore di Bue" combination was decreased of 39%, whereas that of "Costoluto Genovese"/"Natalia" of 11%, compared to control plants. Proteomic results showed a higher representation of proteins associated with pathogen infection in the tolerant rootstock, compared to the sensitive one, meaning a direct involvement of plant defence mechanisms in the tomato response to the pathogen challenge.

Proteomic analysis of apricot fruit during ripening.

Ripening of climacteric fruits involves a complex network of biochemical and metabolic changes that make them palatable and rich in nutritional and health-beneficial compounds. Since fruit maturation has a profound impact on human nutrition, it has been recently the object of increasing research activity by holistic approaches, especially on model species. Here we report on the original proteomic characterization of ripening in apricot, a widely cultivated species of temperate zones appreciated for its taste and aromas, whose cultivation is yet hampered by specific limitations. Fruits of Prunus armeniaca cv. Vesuviana were harvested at three ripening stages and proteins extracted and resolved by 1D and 2D electrophoresis. Whole lanes from 1D gels were subjected to shot-gun analysis that identified 245 gene products, showing preliminary qualitative differences between maturation stages. In parallel, differential analysis of 2D proteomic maps highlighted 106 spots as differentially represented among variably ripen fruits. Most of these were further identified by means of MALDI-TOF-PMF and nanoLC-ESI-LIT-MS/MS as enzymes involved in main biochemical processes influencing metabolic/structural changes occurring during maturation, i.e. organic acids, carbohydrates and energy metabolism, ethylene biosynthesis, cell wall restructuring and stress response, or as protein species linkable to peculiar fruit organoleptic characteristics. In addition to originally present preliminary information on the main biochemical changes that characterize apricot ripening, this study also provides indications for future marker-assisted selection breeding programs aimed to ameliorate fruit quality.

Proteomic analysis of temperature stress-responsive proteins in Arabidopsis thaliana rosette leaves.

Plants, as sessile organisms, are continuously exposed to temperature changes in the environment. Low and high temperature stresses have a great impact on agricultural productivity, since they significantly alter plant metabolism and physiology. Plant response to temperature stress is a quantitative character, being influenced by the degree of stress, time of exposure, as well as plant adaptation ability; it involves profound cellular changes at the proteomic level. We describe here the quantitative variations of the protein repertoire of Arabidopsis thaliana rosette leaves after exposing seedlings to either short-term cold or heat temperature stress. A proteomic approach, based on two-dimensional electrophoresis and MALDI-TOF peptide mass fingerprinting and/or nanoLC-ESI-LIT-MS/MS experiments, was used for this purpose. The comparison of the resulting proteomic maps highlighted proteins showing quantitative variations induced by temperature treatments. Thirty-eight protein spots exhibited significant quantitative changes under at least one stress condition. Identified, differentially-represented proteins belong to two main broad functional groups, namely energy production/carbon metabolism and response to abiotic and oxidative stresses. The role of the identified proteins is discussed here in relation to plant adaptation to cold or heat stresses. Our results suggest a significant overlapping of the responses to opposite temperature extremes.

Lens culinaris Medik. seed proteome: analysis to identify landrace markers.

Unlike modern cultivars selected for their growth performances in specific environmental conditions, local landraces have a high genetic variability that is an important resource for plant breeding. Consequent to their high adaptation to different environmental conditions, these landraces may have evolved adaptive gene complexes To promote the survival of endangered lentil landraces, we previously investigated the genetic relationship between two ancient landraces cultivated in the Molise region (Capracotta and Conca Casale, south-central Italy) and widely spread commercial varieties using an integrated approach consisting of morphological, DNA and protein characterization. In the present study, we used a proteomic approach to compare the mature seed proteomes of the Capracotta and Conca Casale lentil landraces. Multivariate analysis of 145 differentially expressed protein spots demonstrated that 52 proteins are required to discriminate among the two landraces. Therefore, these 52 proteins can be considered "landrace markers". The results of this study show that the combination of proteomics and multivariate analysis can be used to identify physiological and/or environmental markers, and is thus a powerful tool that complements the analysis of biodiversity in plant ecotypes.