Centro Of Ricerca Per La Cerealicoltura

Foggia, Italy

Centro Of Ricerca Per La Cerealicoltura

Foggia, Italy

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Trono D.,Centro Of Ricerca Per La Cerealicoltura | Laus M.N.,University of Foggia | Soccio M.,University of Foggia | Alfarano M.,University of Foggia | Pastore D.,University of Foggia
Frontiers in Plant Science | Year: 2015

In plants, the existence of a mitochondrial potassium channel was firstly demonstrated about 15 years ago in durum wheat as an ATP-dependent potassium channel (PmitoKATP). Since then, both properties of the original PmitoKATP and occurrence of different mitochondrial potassium channels in a number of plant species (monocotyledonous and dicotyledonous) and tissues/organs (etiolated and green) have been shown. Here, an overview of the current knowledge is reported; in particular, the issue of PmitoKATP physiological modulation is addressed. Similarities and differences with other potassium channels, as well as possible cross-regulation with other mitochondrial proteins (Plant Uncoupling Protein, Alternative Oxidase, Plant Inner Membrane Anion Channel) are also described. PmitoKATP is inhibited by ATP and activated by superoxide anion, as well as by free fatty acids (FFAs) and acyl-CoAs. Interestingly, channel activation increases electrophoretic potassium uptake across the inner membrane toward the matrix, so collapsing membrane potential (ΔΨ), the main component of the protonmotive force (Δp) in plant mitochondria; moreover, cooperation between PmitoKATP and the K+/H+ antiporter allows a potassium cycle able to dissipate also ΔpH. Interestingly, ΔΨ collapse matches with an active control of mitochondrial reactive oxygen species (ROS) production. Fully open channel is able to lower superoxide anion up to 35-fold compared to a condition of ATP-inhibited channel. On the other hand, ΔΨ collapse by PmitoKATP was unexpectedly found to not affect ATP synthesis via oxidative phosphorylation. This may probably occur by means of a controlled collapse due to ATP inhibition of PmitoKATP; this brake to the channel activity may allow a loss of the bulk phase Δp, but may preserve a non-classically detectable localized driving force for ATP synthesis. This ability may become crucial under environmental/oxidative stress. In particular, under moderate hyperosmotic stress (mannitol or NaCl), PmitoKATP was found to be activated by ROS, so inhibiting further large-scale ROS production according to a feedback mechanism; moreover, a stress-activated phospholipase A2 may generate FFAs, further activating the channel. In conclusion, a main property of PmitoKATP is the ability to keep in balance the control of harmful ROS with the mitochondrial/cellular bioenergetics, thus preserving ATP for energetic needs of cell defense under stress. © 2015 Trono, Laus, Soccio, Alfarano and Pastore.


Borrelli G.M.,Centro Of Ricerca Per La Cerealicoltura | Trono D.,Centro Of Ricerca Per La Cerealicoltura
International Journal of Molecular Sciences | Year: 2016

Secondary metabolites, also known as phytochemicals, represent a large subset of plant molecules that include compounds with health-promoting effects. Indeed, a number of epidemiological studies have shown that, when taken regularly and in adequate amounts, these molecules can have long-term beneficial effects on human health, through reduction of the incidence of degenerative diseases, such as cardiovascular diseases, obesity, diabetes, and cancer. As the dietary intake of these phytochemicals is often inadequate, various strategies are in use to improve their content in staple crops, and the end-products thereof. One of the most effective strategies is crop improvement through genetic approaches, as this is the only way to generate new cultivars in which the high accumulation of a given phytochemical is stably fixed. Efforts to genetically improve quality traits are rapidly evolving, from classical breeding to molecular-assisted approaches; these require sound understanding of the molecular bases underlying the traits, to identify the genes/alleles that control them. This can be achieved through global analysis of the metabolic pathway responsible for phytochemical accumulation, to identify the link between phytochemical content and the activities of key enzymes that regulate the metabolic pathway, and between the key enzymes and their encoding genes/alleles. Once these have been identified, they can be used as markers for selection of new improved genotypes through biotechnological approaches. This review provides an overview of the major health-promoting properties shown to be associated with the dietary intake of phytochemicals, and describes how molecular approaches provide means for improving the health quality of edible crops. Finally, a case study is illustrated, of the identification in durum wheat of the Lipoxygenase-B1 genes that control the final carotenoid content in semolina-based foods, such as pasta products. © 2016 by the authors; licensee MDPI, Basel, Switzerland.


Canfora L.,Italian Agricultural Research Council | Malusa E.,Italian Agricultural Research Council | Malusa E.,Research Institute of Horticulture | Salvati L.,Italian Agricultural Research Council | And 3 more authors.
Applied Soil Ecology | Year: 2015

This study evaluates the effect of two different kinds of liquid organic fertilizers and a mineral fertilizer on microbial populations (Eubacteria and Archaea) living in the tomato rhizosphere. The organic fertilizers comprised a stillage from bakery yeasts production, characterized by acidic pH and high organic nitrogen content, as well as a vermicompost extract with alkaline pH and low nitrogen content. The number of operational taxonomic units (OTUs) of Archaea species significantly increased after the addition of both the organic and mineral fertilizers, in comparison to the unfertilized control. Eubacteria OTUs increased only in plants treated with the stillage. T-RFLPs profiles identified different clusters of soil rhizospheric Archaea and Eubacteria in treatments of both organic and mineral fertilizers. However, multivariate analyses performed on indicators of genetic diversity, based on the T-RFLPs profiles, and Shannon and equitability indexes of the Archaea and Eubacteria communities, pointed out marked differences between the control (or the mineral fertilization) and the organic fertilizers. The latter treatments induced also a higher root growth and a modification of the root architecture in comparison to both control and mineral fertilizer. The results are discussed with reference to the effect of the organic products on rhizospheric soil characteristics and the relationships between the plant and the rhizospheric microorganisms. © 2014 Elsevier B.V.


Borrelli G.M.,Centro Of Ricerca Per La Cerealicoltura | Trono D.,Centro Of Ricerca Per La Cerealicoltura
International Journal of Molecular Sciences | Year: 2015

Lipases and phospholipases are interfacial enzymes that hydrolyze hydrophobic ester linkages of triacylglycerols and phospholipids, respectively. In addition to their role as esterases, these enzymes catalyze a plethora of other reactions; indeed, lipases also catalyze esterification, transesterification and interesterification reactions, and phospholipases also show acyltransferase, transacylase and transphosphatidylation activities. Thus, lipases and phospholipases represent versatile biocatalysts that are widely used in various industrial applications, such as for biodiesels, food, nutraceuticals, oil degumming and detergents; minor applications also include bioremediation, agriculture, cosmetics, leather and paper industries. These enzymes are ubiquitous in most living organisms, across animals, plants, yeasts, fungi and bacteria. For their greater availability and their ease of production, microbial lipases and phospholipases are preferred to those derived from animals and plants. Nevertheless, traditional purification strategies from microbe cultures have a number of disadvantages, which include non-reproducibility and low yields. Moreover, native microbial enzymes are not always suitable for biocatalytic processes. The development of molecular techniques for the production of recombinant heterologous proteins in a host system has overcome these constraints, as this allows high-level protein expression and production of new redesigned enzymes with improved catalytic properties. These can meet the requirements of specific industrial process better than the native enzymes. The purpose of this review is to give an overview of the structural and functional features of lipases and phospholipases, to describe the recent advances in optimization of the production of recombinant lipases and phospholipases, and to summarize the information available relating to their major applications in industrial processes. © 2015 by the authors; licensee MDPI, Basel, Switzerland.


Garofalo P.,Unita di Ricerca per i Sistemi Colturali degli Ambienti caldoaridi | Rinaldi M.,Unita di Ricerca per i Sistemi Colturali degli Ambienti caldoaridi | Rinaldi M.,Centro Of Ricerca Per La Cerealicoltura
Italian Journal of Agrometeorology | Year: 2013

In water-limited environments, water is the main limiting factor of crop production, especially in rainfed crops such as durum winter wheat (Triticum durum Desf.). Consequently, also in climatic change projections, it is essential both to recognise characteristics in breeding programs that can lead to drought tolerance and to reduce the time needed to observe variations of these traits on crop yield. Moreover, changing in management strategies could improve crop adaptation to climate change, not considered in this approach. Crop growth models can assist breeding research in identifying these traits. The CropSyst model was parameterized for durum wheat cultivated in Southern Italy: crop characteristics were analyzed, development (grain filling duration and phenologic response to water stress), canopy expansion (specific leaf area, leaf duration and ratio between leaf and stem) and water uptake (root length). Model sensitivity was evaluated varying one parameter at a time and changing the value by ± 5, ± 10 and ± 20% of calibrated values. Wheat was simulated with past real daily climatic data (55 years, from 1952 to 2006) and future daily climatic data predicted with an HADCM3 global climatic model (100 years, from 2000 to 2100) where an average air temperature increase of +2 °C is expected as well as a CO2 concentration of 550 ppm (IPCC, A2 scenario). Leaf area duration and specific leaf area proved to be the parameters with the greatest impacts on wheat yield, with changing in wheat yield from -20 to16% and from -38 to 35% respectively as consequence to variation for these parameters oscillating between - 20 and 20%. The ratio between the leaf and the stem biomass accumulation was inversely and linearly related to grain yield. Lengthening or shortening the grain filling duration did not seem to provide benefits in term of grain yield. The "non-response" in term of grain yield to water stress highlights that the wheat crop was optimized yet in Mediterranean environment in order to maintain production stability in drought conditions which could accelerate different crop phenological stages. The variation of maximum root depth (from 0.8 to 1.2 m) did not result in any significant variation in grain yield. The changes of crop morphology could also enhance climate change adaptation.


Mazzucotelli E.,Centro Of Ricerca Per La Genomica Vegetale | Trono D.,Centro Of Ricerca Per La Cerealicoltura
Plant Science | Year: 2015

We previously isolated four cDNAs in durum wheat, TdsPLA2I, TdsPLA2II, TdsPLA2III and TdsPLA2IV, that encode proteins with homology to plant secretory phospholipases A2 (sPLA2s) (Verlotta et al., Int. J. Mol. Sci., 14, 2013, 5146-5169). In this study, we have further characterized TdsPLA2II and TdsPLA2III sequences that, on the basis of our previous findings, might encode sPLA2 isoforms with different features. Functional analysis revealed that, similarly to other known sPLA2s, TdsPLA2II and TdsPLA2III have an optimum at pH 9.0, require Ca2+, are heat stable, and are inhibited by the disulfide-bond-reducing agent dithiothreitol. However, differences emerged between these TdsPLA2 isoforms. Transcript analysis revealed that the TdsPLA2III gene is highly up-regulated under different environmental stresses; conversely, the TdsPLA2II gene is expressed at constant levels under almost all of the stress conditions examined. Moreover, TdsPLA2II is saturated at micromolar substrate and Ca2+ concentrations, whereas TdsPLA2III requires millimolar concentrations to reach maximal activity. This suggests that TdsPLA2II normally functions under optimal conditions in vivo, whereas TdsPLA2III is only partially activated, depending on the specific phospholipid and Ca2+ levels. Altogether these data lead to the hypothesis that in vivo TdsPLA2II and TdsPLA2III are differently regulated at both molecular and biochemical level and that TdsPLA2III plays a major role in durum wheat response to adverse environmental conditions. © 2015 Elsevier Ireland Ltd.


PubMed | University of Foggia and Centro Of Ricerca Per La Cerealicoltura
Type: | Journal: Frontiers in plant science | Year: 2015

In plants, the existence of a mitochondrial potassium channel was firstly demonstrated about 15 years ago in durum wheat as an ATP-dependent potassium channel (PmitoKATP). Since then, both properties of the original PmitoKATP and occurrence of different mitochondrial potassium channels in a number of plant species (monocotyledonous and dicotyledonous) and tissues/organs (etiolated and green) have been shown. Here, an overview of the current knowledge is reported; in particular, the issue of PmitoKATP physiological modulation is addressed. Similarities and differences with other potassium channels, as well as possible cross-regulation with other mitochondrial proteins (Plant Uncoupling Protein, Alternative Oxidase, Plant Inner Membrane Anion Channel) are also described. PmitoKATP is inhibited by ATP and activated by superoxide anion, as well as by free fatty acids (FFAs) and acyl-CoAs. Interestingly, channel activation increases electrophoretic potassium uptake across the inner membrane toward the matrix, so collapsing membrane potential (), the main component of the protonmotive force (p) in plant mitochondria; moreover, cooperation between PmitoKATP and the K(+)/H(+) antiporter allows a potassium cycle able to dissipate also pH. Interestingly, collapse matches with an active control of mitochondrial reactive oxygen species (ROS) production. Fully open channel is able to lower superoxide anion up to 35-fold compared to a condition of ATP-inhibited channel. On the other hand, collapse by PmitoKATP was unexpectedly found to not affect ATP synthesis via oxidative phosphorylation. This may probably occur by means of a controlled collapse due to ATP inhibition of PmitoKATP; this brake to the channel activity may allow a loss of the bulk phase p, but may preserve a non-classically detectable localized driving force for ATP synthesis. This ability may become crucial under environmental/oxidative stress. In particular, under moderate hyperosmotic stress (mannitol or NaCl), PmitoKATP was found to be activated by ROS, so inhibiting further large-scale ROS production according to a feedback mechanism; moreover, a stress-activated phospholipase A2 may generate FFAs, further activating the channel. In conclusion, a main property of PmitoKATP is the ability to keep in balance the control of harmful ROS with the mitochondrial/cellular bioenergetics, thus preserving ATP for energetic needs of cell defense under stress.


PubMed | Centro Of Ricerca Per La Cerealicoltura
Type: Journal Article | Journal: International journal of molecular sciences | Year: 2015

Lipases and phospholipases are interfacial enzymes that hydrolyze hydrophobic ester linkages of triacylglycerols and phospholipids, respectively. In addition to their role as esterases, these enzymes catalyze a plethora of other reactions; indeed, lipases also catalyze esterification, transesterification and interesterification reactions, and phospholipases also show acyltransferase, transacylase and transphosphatidylation activities. Thus, lipases and phospholipases represent versatile biocatalysts that are widely used in various industrial applications, such as for biodiesels, food, nutraceuticals, oil degumming and detergents; minor applications also include bioremediation, agriculture, cosmetics, leather and paper industries. These enzymes are ubiquitous in most living organisms, across animals, plants, yeasts, fungi and bacteria. For their greater availability and their ease of production, microbial lipases and phospholipases are preferred to those derived from animals and plants. Nevertheless, traditional purification strategies from microbe cultures have a number of disadvantages, which include non-reproducibility and low yields. Moreover, native microbial enzymes are not always suitable for biocatalytic processes. The development of molecular techniques for the production of recombinant heterologous proteins in a host system has overcome these constraints, as this allows high-level protein expression and production of new redesigned enzymes with improved catalytic properties. These can meet the requirements of specific industrial process better than the native enzymes. The purpose of this review is to give an overview of the structural and functional features of lipases and phospholipases, to describe the recent advances in optimization of the production of recombinant lipases and phospholipases, and to summarize the information available relating to their major applications in industrial processes.


Secondary metabolites, also known as phytochemicals, represent a large subset of plant molecules that include compounds with health-promoting effects. Indeed, a number of epidemiological studies have shown that, when taken regularly and in adequate amounts, these molecules can have long-term beneficial effects on human health, through reduction of the incidence of degenerative diseases, such as cardiovascular diseases, obesity, diabetes, and cancer. As the dietary intake of these phytochemicals is often inadequate, various strategies are in use to improve their content in staple crops, and the end-products thereof. One of the most effective strategies is crop improvement through genetic approaches, as this is the only way to generate new cultivars in which the high accumulation of a given phytochemical is stably fixed. Efforts to genetically improve quality traits are rapidly evolving, from classical breeding to molecular-assisted approaches; these require sound understanding of the molecular bases underlying the traits, to identify the genes/alleles that control them. This can be achieved through global analysis of the metabolic pathway responsible for phytochemical accumulation, to identify the link between phytochemical content and the activities of key enzymes that regulate the metabolic pathway, and between the key enzymes and their encoding genes/alleles. Once these have been identified, they can be used as markers for selection of new improved genotypes through biotechnological approaches. This review provides an overview of the major health-promoting properties shown to be associated with the dietary intake of phytochemicals, and describes how molecular approaches provide means for improving the health quality of edible crops. Finally, a case study is illustrated, of the identification in durum wheat of the Lipoxygenase-B1 genes that control the final carotenoid content in semolina-based foods, such as pasta products.


PubMed | Centro Of Ricerca Per La Genomica Vegetale and Centro Of Ricerca Per La Cerealicoltura
Type: | Journal: Plant science : an international journal of experimental plant biology | Year: 2015

We previously isolated four cDNAs in durum wheat, TdsPLA2I, TdsPLA2II, TdsPLA2III and TdsPLA2IV, that encode proteins with homology to plant secretory phospholipases A2 (sPLA2s) (Verlotta et al., Int. J. Mol. Sci., 14, 2013, 5146-5169). In this study, we have further characterized TdsPLA2II and TdsPLA2III sequences that, on the basis of our previous findings, might encode sPLA2 isoforms with different features. Functional analysis revealed that, similarly to other known sPLA2s, TdsPLA2II and TdsPLA2III have an optimum at pH 9.0, require Ca(2+), are heat stable, and are inhibited by the disulfide-bond-reducing agent dithiothreitol. However, differences emerged between these TdsPLA2 isoforms. Transcript analysis revealed that the TdsPLA2III gene is highly up-regulated under different environmental stresses; conversely, the TdsPLA2II gene is expressed at constant levels under almost all of the stress conditions examined. Moreover, TdsPLA2II is saturated at micromolar substrate and Ca(2+) concentrations, whereas TdsPLA2III requires millimolar concentrations to reach maximal activity. This suggests that TdsPLA2II normally functions under optimal conditions in vivo, whereas TdsPLA2III is only partially activated, depending on the specific phospholipid and Ca(2+) levels. Altogether these data lead to the hypothesis that in vivo TdsPLA2II and TdsPLA2III are differently regulated at both molecular and biochemical level and that TdsPLA2III plays a major role in durum wheat response to adverse environmental conditions.

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