Plant defense against pests and pathogens is known to be conferred by either salicylic acid (SA) ... more Plant defense against pests and pathogens is known to be conferred by either salicylic acid (SA) or jasmonic acid (JA)/ethylene (ET) pathways, depending on infection or herbivore-grazing strategy. It is well attested that SA and JA/ET pathways are mutually antagonistic allowing defense responses to be tailored to particular biotic stresses. Nitric oxide (NO) has emerged as a major signal influencing resistance mediated by both signaling pathways but no attempt has been made to integrate NO into established SA/JA/ET interactions. NO has been shown to act as an inducer or suppressor of signaling along each pathway. NO will initiate SA biosynthesis and nitrosylate key cysteines on TGA-class transcription factors to aid in the initiation of SA-dependent gene expression. Against this, S-nitrosylation of NONEXPRESSOR OF PATHOGENESIS-RELATED PROTEINS1 (NPR1) will promote the NPR1 oligomerization within the cytoplasm to reduce TGA activation. In JA biosynthesis, NO will initiate the expression of JA biosynthetic enzymes, presumably to overcome any antagonistic effects of SA on JA-mediated transcription. NO will also initiate the expression of ET biosynthetic genes but a suppressive role is also observed in the S-nitrosylation and inhibition of S-adenosylmethionine transferases which provides methyl groups for ET production. Based on these data a model for NO action is proposed but we have also highlighted the need to understand when and how inductive and suppressive steps are used.
Thermoplastic, polysaccharide-based plastics are environmentally friendly. However, typical short... more Thermoplastic, polysaccharide-based plastics are environmentally friendly. However, typical shortcomings include lack of water resistance and poor mechanical properties. Nanocomposite manufacturing using pure, highly linear, polysaccharides can overcome such limitations. Cast nanocomposites were fabricated with plant engineered pure amylose (AM), produced in bulk quantity in transgenic barley grain, and cellulose nanofibers (CNF), extracted from agrowaste sugar beet pulp. Morphology, crystallinity, chemical heterogeneity, mechanics, dynamic mechanical, gas and water permeability, and contact angle of the films were investigated. Blending CNF into the AM matrix significantly enhanced the crystallinity, mechanical properties and permeability, whereas glycerol increased elongation at break, mainly by plasticizing the AM. There was significant phase separation between AM and CNF. Dynamic plasticizing and anti-plasticizing effects of both CNF and glycerol were demonstrated by NMR demonstrating high molecular order, but also non-crystalline, and evenly distributed 20 nm-sized glycerol domains. This study demonstrates a new lead in functional polysaccharide-based bioplastic systems.
Background: Sample preparation is a critical process for proteomic studies. Many efficient and re... more Background: Sample preparation is a critical process for proteomic studies. Many efficient and reproducible sample preparation methods have been developed for mass spectrometry-based proteomic analysis of human and animal tissues or cells, but no attempt has been made to evaluate these protocols for plants. We here present an LC-MS/ MS-based proteomics study of barley leaf aimed at optimization of methods to achieve efficient and unbiased trypsin digestion of proteins prior to LC-MS/MS based sequencing and quantification of peptides. We evaluated two spin filter-aided sample preparation protocols using either sodium dodecyl-sulphate or sodium deoxycholate (SDC), and three in-solution digestion (ISD) protocols using SDC or trichloroacetic acid/acetone precipitation. Results: The proteomics workflow identified and quantified up to 1800 barley proteins based on sequencing of up to 6900 peptides per sample. The two spin filter-based protocols provided a 12-38% higher efficiency than the ISD protocols, including more proteins of low abundance. Among the ISD protocols, a simple one-step reduction and S-alkylation method (OP-ISD) was the most efficient for barley leaf sample preparation; it identified and quantified 1500 proteins and displayed higher peptide-to-protein inference ratio and higher average amino acid sequence coverage of proteins. The two spin filter-aided sample preparation protocols are compatible with TMT labelling for quantitative proteomics studies. They exhibited complementary performance as about 30% of the proteins were identified by either one or the other protocol, but also demonstrated a positive bias for membrane proteins when using SDC as detergent. Conclusions: We provide detailed protocols for efficient plant protein sample preparation for LC-MS/MS-based proteomics studies. Spin filter-based protocols are the most efficient for the preparation of leaf samples for MS-based proteomics. However, a simple protocol provides comparable results although with different peptide digestion profile.
Timely perception of adverse environmental changes is critical for survival. Dynamic changes in g... more Timely perception of adverse environmental changes is critical for survival. Dynamic changes in gases are important cues for plants to sense environmental perturbations, such as submergence. In Arabidopsis thaliana, changes in oxygen and nitric oxide (NO) control the stability of ERFVII transcription factors. ERFVII proteolysis is regulated by the N-degron pathway and mediates adaptation to flooding-induced hypoxia. However, how plants detect and transduce early submergence signals remains elusive. Here we show that plants can rapidly detect submergence through passive ethylene entrapment and use this signal to pre-adapt to impending hypoxia. Ethylene can enhance ERFVII stability prior to hypoxia by increasing the NO-scavenger PHYTOGLOBIN1. This ethylene-mediated NO depletion and consequent ERFVII accumulation pre-adapts plants to survive subsequent hypoxia. Our results reveal the biological link between three gaseous signals for the regulation of flooding survival and identifies no...
Symbiotic hemoglobins provide O2 to N2 -fixing bacteria within legume nodules, but the functions ... more Symbiotic hemoglobins provide O2 to N2 -fixing bacteria within legume nodules, but the functions of nonsymbiotic hemoglobins or phytoglobins (Glbs) are much less defined. Immunolabeling combined with confocal microscopy of the Glbs tagged at the C-terminus with green fluorescent protein was used to determine their subcellular localizations in Arabidopsis and Lotus japonicus. Recombinant proteins were used to examine NO scavenging in vitro and transgenic plants to show S-nitrosylation and other in vivo interactions with NO and ABA responses. We found that Glbs occur in the nuclei, chloroplasts, and amyloplasts of both model plants, and also in the cytoplasm of Arabidopsis cells. The proteins show similar NO dioxygenase activities in vitro, are nitrosylated in Cys residues in vivo, and scavenge NO in the stomatal cells. The Cys/Ser mutation does not affect NO dioxygenase activity, and S-nitrosylation does not significantly consume NO. We demonstrate an interaction between Glbs and ABA on several grounds: Glb1 and Glb2 scavenge NO produced in stomatal guard cells following ABA supply; plants overexpressing Glb1 show higher constitutive expression of the ABA responsive genes Responsive to ABA (RAB18), Responsive to Dehydration (RD29A), and Highly ABA-Induced 2 (HAI2), and are more tolerant to dehydration; and ABA strongly up-regulates class 1 Glbs. We conclude that Glbs modulate NO and interact with ABA in crucial physiological processes such as the plant's response to dessication. This article is protected by copyright. All rights reserved.
meta-Tyrosine (m-Tyr) is a non-protein amino acid produced in both plants and animals. Primary mo... more meta-Tyrosine (m-Tyr) is a non-protein amino acid produced in both plants and animals. Primary mode of action of this phenylalanine analog is its incorporation into protein structure leading to formation of aberrant molecules. An increased level of m-Tyr in animal cells is detected under oxidative stress and during age-related processes characterized by overproduction of reactive oxygen species (ROS). The aim of this study was to link m-Tyr physiological action to disturbances in ROS metabolism in tomato (Solanum lycopersicum L.) seedlings roots. Treatment of tomato seedlings with m-Tyr (50 or 250 μM) for 24-72 h led to inhibition of root growth without a lethal effect. Toxicity of m-Tyr after 72 h was connected with an increase in hydrogen peroxide concentration in roots and ROS leakage into the surrounding medium. On the contrary, membrane permeability and lipid peroxidation in roots were the same as for the control. This was accompanied by a decrease in total antioxidant activity...
Plant science : an international journal of experimental plant biology, 2017
Class 1 plant hemoglobins (phytoglobins) are upregulated during low-oxygen stress and participate... more Class 1 plant hemoglobins (phytoglobins) are upregulated during low-oxygen stress and participate in metabolism and cell signaling via modulation of the levels of nitric oxide (NO). We studied the effects of overexpression and knockdown of the class 1 phytoglobin gene in barley (Hordeum vulgare L.) under low-oxygen stress. The overexpression of phytoglobin reduced the amount of NO released, while knockdown significantly stimulated NO emission. It has previously been shown that NO inhibits aconitase activity, so decreased aconitase activity in knockdown plants acts as a biomarker for high internal NO levels. The overexpression of phytoglobin corresponded to higher ATP/ADP ratios, pyrophosphate levels and aconitase activity under anoxia, while knockdown of phytoglobin resulted in the increased level of protein nitrosylation, elevation of alcohol dehydrogenase and nitrosoglutathione reductase activities. The overexpressing plants showed various signs of stunted growth under normoxia, b...
Nitric oxide (NO) is a key messenger in plant stress responses but its exact role in drought resp... more Nitric oxide (NO) is a key messenger in plant stress responses but its exact role in drought response remains unclear. To investigate the role of NO in drought response we employed transgenic barley plants (UHb) overexpressing the barley non-symbiotic hemoglobin gene HvHb1 that oxidizes NO to NO3-. Reduced NO production under drought conditions in UHb plants was associated with increased drought tolerance. Since NO biosynthesis has been related to polyamine metabolism, we investigated whether the observed drought-related NO changes could involve polyamine pathway. UHb plants showed increases in total polyamines and in particular polyamines such as spermidine. These increases correlated with the accumulation of the amino acid precursors of polyamines and with the expression of specific polyamine biosynthesis genes. This suggests a potential interplay between NO and polyamine biosynthesis during drought response. Since ethylene has been linked to NO signaling and it is also related to...
International journal of molecular sciences, Jan 30, 2017
Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solu... more Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solutions to solve immediate environmental issues. Polysaccharide-based bio-plastics represent important alternatives to conventional plastic because of their intrinsic biodegradable nature. Amylose-only (AO), an engineered barley starch with 99% amylose, was tested to produce cross-linked all-natural bioplastic using normal barley starch as a control. Glycerol was used as plasticizer and citrate cross-linking was used to improve the mechanical properties of cross-linked AO starch extrudates. Extrusion converted the control starch from A-type to Vh- and B-type crystals, showing a complete melting of the starch crystals in the raw starch granules. The cross-linked AO and control starch specimens displayed an additional wide-angle diffraction reflection. Phospholipids complexed with Vh-type single helices constituted an integrated part of the AO starch specimens. Gas permeability tests of sele...
Starch phosphorylation occurs naturally during starch metabolism in the plant and is catalysed by... more Starch phosphorylation occurs naturally during starch metabolism in the plant and is catalysed by glucan water dikinases (GWD1) and phosphoglucan water dikinase/glucan water dikinase 3 (PWD/GWD3). We generated six stable individual transgenic lines by over-expressing the potato GWD1 in rice. Transgenic rice grain starch had 9-fold higher 6-phospho (6-P) monoesters and double amounts of 3-phospho (3-P) monoesters, respectively, compared to control grain. The shape and topography of the transgenic starch granules were moderately altered including surface pores and less well defined edges. The gelatinization temperatures of both rice flour and extracted starch were significantly lower than those of the control and hence negatively correlated with the starch phosphate content. The 6-P content was positively correlated with amylose content and relatively long amylopectin chains with DP25-36, and the 3-P content was positively correlated with short chains of DP6-12. The starch pasting tem...
International journal of biological macromolecules, Jan 29, 2017
This study investigated the influence of diurnal photosynthetic activity on the morphology, molec... more This study investigated the influence of diurnal photosynthetic activity on the morphology, molecular composition, crystallinity, and gelatinization properties of normal barley starch (NBS) and waxy barley starch (WBS) granules from plants cultivated in a greenhouse under normal diurnal (16h light) or constant light photosynthetic conditions. Growth rings were observed in all starch samples regardless of lighting conditions. The size distribution of whole and debranched WBS analyzed by gel-permeation chromatography did not appear to be influenced by the different lighting regimes, however, a greater relative crystallinity measured by wide-angle X-ray scattering and greater crystalline quality as judged by differential scanning calorimetry was observed under the diurnal lighting regime. NBS cultivated under the diurnal photosynthetic lighting regime displayed lower amylose content (18.7%), and shorter amylose chains than its counterpart grown under constant light. Although the relati...
Plant science : an international journal of experimental plant biology, 2017
Grasses such as wheat and barley constitute some of the main crops currently cultivated worldwide... more Grasses such as wheat and barley constitute some of the main crops currently cultivated worldwide. Cultivation of wild cereals such as Hordeum spontaneum (wild barley) and Triticum diccocoides (wild emmer) started in southwest Asia. Despite the many studies on the origins of agriculture and plant domestication, surprisingly few studies have discussed the importance of the nutritional quality of barley and emmer wheat in their evolution. A comparison of domesticated forms of these cereals with their wild progenitors suggests an evolution towards bigger grains with higher glycemic loads in the form of higher relative starch content with lower relative protein, fiber and mineral contents. In this work we hypothesize that in addition to the simple explanation that larger grains emerged under domestication, complex and indirect effects such as increased glycemic index and sweet taste should also be taken into consideration.
The plant-specific NAC transcription factors have attracted particular attention because of their... more The plant-specific NAC transcription factors have attracted particular attention because of their involvement in stress responses, senescence, and nutrient remobilization. The HvNAC005 gene of barley encodes a protein belonging to subgroup NAC-a6 of the NAC family. This study shows that HvNAC005 is associated with developmental senescence. It was significantly up-regulated following ABA treatment, supported by ABA-responsive elements in its promoter, but it was not up-regulated during dark-induced senescence. The C-termini of proteins closely related to HvNAC005 showed overall high divergence but also contained conserved short motifs. A serine- and leucine-containing central motif was essential for transcriptional activity of the HvNAC005 C-terminus in yeast. Over-expression of HvNAC005 in barley resulted in a strong phenotype with delayed development combined with precocious senescence. The over-expressing plants showed up-regulation of genes involved with secondary metabolism, hor...
Users may download and print one copy of any publication from the public portal for the purpose... more Users may download and print one copy of any publication from the public portal for the purpose of private study or research. You may not further distribute the material or use it for any profit-making activity or commercial gain You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.
International journal of biological macromolecules, 2016
The effects of amylose deposition on crystalline regions of barley starch granules were studied i... more The effects of amylose deposition on crystalline regions of barley starch granules were studied in granules containing zero to 99.1% amylose using "waxy" (WBS, 0% amylose), normal (NBS, 18% amylose) and amylose-only barley lines (AOS, 99.1% amylose). The effects were probed after hydrolysis of amorphous regions of starch granules in dilute HCl generating lintners, which typically represent the crystalline lamella of starch granules. Compared to NBS and WBS, AOS granules exhibited an irregular, multilobular morphology with a rough surface texture. AOS displayed lower rates of acid hydrolysis than WBS, and AOS reached a plateau at ∼45wt% acid hydrolysis. High-performance anion-exchange chromatography of lintners at equivalent levels of hydrolysis (45wt%) revealed the average degree of polymerization (DP) of AOS lintners was 21, substantially smaller than that of NBS and WBS (DP 42). AOS lintners contained the lowest number of chains (NC) per molecule (1.1) compared to NBS (2...
Plant defense against pests and pathogens is known to be conferred by either salicylic acid (SA) ... more Plant defense against pests and pathogens is known to be conferred by either salicylic acid (SA) or jasmonic acid (JA)/ethylene (ET) pathways, depending on infection or herbivore-grazing strategy. It is well attested that SA and JA/ET pathways are mutually antagonistic allowing defense responses to be tailored to particular biotic stresses. Nitric oxide (NO) has emerged as a major signal influencing resistance mediated by both signaling pathways but no attempt has been made to integrate NO into established SA/JA/ET interactions. NO has been shown to act as an inducer or suppressor of signaling along each pathway. NO will initiate SA biosynthesis and nitrosylate key cysteines on TGA-class transcription factors to aid in the initiation of SA-dependent gene expression. Against this, S-nitrosylation of NONEXPRESSOR OF PATHOGENESIS-RELATED PROTEINS1 (NPR1) will promote the NPR1 oligomerization within the cytoplasm to reduce TGA activation. In JA biosynthesis, NO will initiate the expression of JA biosynthetic enzymes, presumably to overcome any antagonistic effects of SA on JA-mediated transcription. NO will also initiate the expression of ET biosynthetic genes but a suppressive role is also observed in the S-nitrosylation and inhibition of S-adenosylmethionine transferases which provides methyl groups for ET production. Based on these data a model for NO action is proposed but we have also highlighted the need to understand when and how inductive and suppressive steps are used.
Thermoplastic, polysaccharide-based plastics are environmentally friendly. However, typical short... more Thermoplastic, polysaccharide-based plastics are environmentally friendly. However, typical shortcomings include lack of water resistance and poor mechanical properties. Nanocomposite manufacturing using pure, highly linear, polysaccharides can overcome such limitations. Cast nanocomposites were fabricated with plant engineered pure amylose (AM), produced in bulk quantity in transgenic barley grain, and cellulose nanofibers (CNF), extracted from agrowaste sugar beet pulp. Morphology, crystallinity, chemical heterogeneity, mechanics, dynamic mechanical, gas and water permeability, and contact angle of the films were investigated. Blending CNF into the AM matrix significantly enhanced the crystallinity, mechanical properties and permeability, whereas glycerol increased elongation at break, mainly by plasticizing the AM. There was significant phase separation between AM and CNF. Dynamic plasticizing and anti-plasticizing effects of both CNF and glycerol were demonstrated by NMR demonstrating high molecular order, but also non-crystalline, and evenly distributed 20 nm-sized glycerol domains. This study demonstrates a new lead in functional polysaccharide-based bioplastic systems.
Background: Sample preparation is a critical process for proteomic studies. Many efficient and re... more Background: Sample preparation is a critical process for proteomic studies. Many efficient and reproducible sample preparation methods have been developed for mass spectrometry-based proteomic analysis of human and animal tissues or cells, but no attempt has been made to evaluate these protocols for plants. We here present an LC-MS/ MS-based proteomics study of barley leaf aimed at optimization of methods to achieve efficient and unbiased trypsin digestion of proteins prior to LC-MS/MS based sequencing and quantification of peptides. We evaluated two spin filter-aided sample preparation protocols using either sodium dodecyl-sulphate or sodium deoxycholate (SDC), and three in-solution digestion (ISD) protocols using SDC or trichloroacetic acid/acetone precipitation. Results: The proteomics workflow identified and quantified up to 1800 barley proteins based on sequencing of up to 6900 peptides per sample. The two spin filter-based protocols provided a 12-38% higher efficiency than the ISD protocols, including more proteins of low abundance. Among the ISD protocols, a simple one-step reduction and S-alkylation method (OP-ISD) was the most efficient for barley leaf sample preparation; it identified and quantified 1500 proteins and displayed higher peptide-to-protein inference ratio and higher average amino acid sequence coverage of proteins. The two spin filter-aided sample preparation protocols are compatible with TMT labelling for quantitative proteomics studies. They exhibited complementary performance as about 30% of the proteins were identified by either one or the other protocol, but also demonstrated a positive bias for membrane proteins when using SDC as detergent. Conclusions: We provide detailed protocols for efficient plant protein sample preparation for LC-MS/MS-based proteomics studies. Spin filter-based protocols are the most efficient for the preparation of leaf samples for MS-based proteomics. However, a simple protocol provides comparable results although with different peptide digestion profile.
Timely perception of adverse environmental changes is critical for survival. Dynamic changes in g... more Timely perception of adverse environmental changes is critical for survival. Dynamic changes in gases are important cues for plants to sense environmental perturbations, such as submergence. In Arabidopsis thaliana, changes in oxygen and nitric oxide (NO) control the stability of ERFVII transcription factors. ERFVII proteolysis is regulated by the N-degron pathway and mediates adaptation to flooding-induced hypoxia. However, how plants detect and transduce early submergence signals remains elusive. Here we show that plants can rapidly detect submergence through passive ethylene entrapment and use this signal to pre-adapt to impending hypoxia. Ethylene can enhance ERFVII stability prior to hypoxia by increasing the NO-scavenger PHYTOGLOBIN1. This ethylene-mediated NO depletion and consequent ERFVII accumulation pre-adapts plants to survive subsequent hypoxia. Our results reveal the biological link between three gaseous signals for the regulation of flooding survival and identifies no...
Symbiotic hemoglobins provide O2 to N2 -fixing bacteria within legume nodules, but the functions ... more Symbiotic hemoglobins provide O2 to N2 -fixing bacteria within legume nodules, but the functions of nonsymbiotic hemoglobins or phytoglobins (Glbs) are much less defined. Immunolabeling combined with confocal microscopy of the Glbs tagged at the C-terminus with green fluorescent protein was used to determine their subcellular localizations in Arabidopsis and Lotus japonicus. Recombinant proteins were used to examine NO scavenging in vitro and transgenic plants to show S-nitrosylation and other in vivo interactions with NO and ABA responses. We found that Glbs occur in the nuclei, chloroplasts, and amyloplasts of both model plants, and also in the cytoplasm of Arabidopsis cells. The proteins show similar NO dioxygenase activities in vitro, are nitrosylated in Cys residues in vivo, and scavenge NO in the stomatal cells. The Cys/Ser mutation does not affect NO dioxygenase activity, and S-nitrosylation does not significantly consume NO. We demonstrate an interaction between Glbs and ABA on several grounds: Glb1 and Glb2 scavenge NO produced in stomatal guard cells following ABA supply; plants overexpressing Glb1 show higher constitutive expression of the ABA responsive genes Responsive to ABA (RAB18), Responsive to Dehydration (RD29A), and Highly ABA-Induced 2 (HAI2), and are more tolerant to dehydration; and ABA strongly up-regulates class 1 Glbs. We conclude that Glbs modulate NO and interact with ABA in crucial physiological processes such as the plant's response to dessication. This article is protected by copyright. All rights reserved.
meta-Tyrosine (m-Tyr) is a non-protein amino acid produced in both plants and animals. Primary mo... more meta-Tyrosine (m-Tyr) is a non-protein amino acid produced in both plants and animals. Primary mode of action of this phenylalanine analog is its incorporation into protein structure leading to formation of aberrant molecules. An increased level of m-Tyr in animal cells is detected under oxidative stress and during age-related processes characterized by overproduction of reactive oxygen species (ROS). The aim of this study was to link m-Tyr physiological action to disturbances in ROS metabolism in tomato (Solanum lycopersicum L.) seedlings roots. Treatment of tomato seedlings with m-Tyr (50 or 250 μM) for 24-72 h led to inhibition of root growth without a lethal effect. Toxicity of m-Tyr after 72 h was connected with an increase in hydrogen peroxide concentration in roots and ROS leakage into the surrounding medium. On the contrary, membrane permeability and lipid peroxidation in roots were the same as for the control. This was accompanied by a decrease in total antioxidant activity...
Plant science : an international journal of experimental plant biology, 2017
Class 1 plant hemoglobins (phytoglobins) are upregulated during low-oxygen stress and participate... more Class 1 plant hemoglobins (phytoglobins) are upregulated during low-oxygen stress and participate in metabolism and cell signaling via modulation of the levels of nitric oxide (NO). We studied the effects of overexpression and knockdown of the class 1 phytoglobin gene in barley (Hordeum vulgare L.) under low-oxygen stress. The overexpression of phytoglobin reduced the amount of NO released, while knockdown significantly stimulated NO emission. It has previously been shown that NO inhibits aconitase activity, so decreased aconitase activity in knockdown plants acts as a biomarker for high internal NO levels. The overexpression of phytoglobin corresponded to higher ATP/ADP ratios, pyrophosphate levels and aconitase activity under anoxia, while knockdown of phytoglobin resulted in the increased level of protein nitrosylation, elevation of alcohol dehydrogenase and nitrosoglutathione reductase activities. The overexpressing plants showed various signs of stunted growth under normoxia, b...
Nitric oxide (NO) is a key messenger in plant stress responses but its exact role in drought resp... more Nitric oxide (NO) is a key messenger in plant stress responses but its exact role in drought response remains unclear. To investigate the role of NO in drought response we employed transgenic barley plants (UHb) overexpressing the barley non-symbiotic hemoglobin gene HvHb1 that oxidizes NO to NO3-. Reduced NO production under drought conditions in UHb plants was associated with increased drought tolerance. Since NO biosynthesis has been related to polyamine metabolism, we investigated whether the observed drought-related NO changes could involve polyamine pathway. UHb plants showed increases in total polyamines and in particular polyamines such as spermidine. These increases correlated with the accumulation of the amino acid precursors of polyamines and with the expression of specific polyamine biosynthesis genes. This suggests a potential interplay between NO and polyamine biosynthesis during drought response. Since ethylene has been linked to NO signaling and it is also related to...
International journal of molecular sciences, Jan 30, 2017
Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solu... more Bio-plastics and bio-materials are composed of natural or biomass derived polymers, offering solutions to solve immediate environmental issues. Polysaccharide-based bio-plastics represent important alternatives to conventional plastic because of their intrinsic biodegradable nature. Amylose-only (AO), an engineered barley starch with 99% amylose, was tested to produce cross-linked all-natural bioplastic using normal barley starch as a control. Glycerol was used as plasticizer and citrate cross-linking was used to improve the mechanical properties of cross-linked AO starch extrudates. Extrusion converted the control starch from A-type to Vh- and B-type crystals, showing a complete melting of the starch crystals in the raw starch granules. The cross-linked AO and control starch specimens displayed an additional wide-angle diffraction reflection. Phospholipids complexed with Vh-type single helices constituted an integrated part of the AO starch specimens. Gas permeability tests of sele...
Starch phosphorylation occurs naturally during starch metabolism in the plant and is catalysed by... more Starch phosphorylation occurs naturally during starch metabolism in the plant and is catalysed by glucan water dikinases (GWD1) and phosphoglucan water dikinase/glucan water dikinase 3 (PWD/GWD3). We generated six stable individual transgenic lines by over-expressing the potato GWD1 in rice. Transgenic rice grain starch had 9-fold higher 6-phospho (6-P) monoesters and double amounts of 3-phospho (3-P) monoesters, respectively, compared to control grain. The shape and topography of the transgenic starch granules were moderately altered including surface pores and less well defined edges. The gelatinization temperatures of both rice flour and extracted starch were significantly lower than those of the control and hence negatively correlated with the starch phosphate content. The 6-P content was positively correlated with amylose content and relatively long amylopectin chains with DP25-36, and the 3-P content was positively correlated with short chains of DP6-12. The starch pasting tem...
International journal of biological macromolecules, Jan 29, 2017
This study investigated the influence of diurnal photosynthetic activity on the morphology, molec... more This study investigated the influence of diurnal photosynthetic activity on the morphology, molecular composition, crystallinity, and gelatinization properties of normal barley starch (NBS) and waxy barley starch (WBS) granules from plants cultivated in a greenhouse under normal diurnal (16h light) or constant light photosynthetic conditions. Growth rings were observed in all starch samples regardless of lighting conditions. The size distribution of whole and debranched WBS analyzed by gel-permeation chromatography did not appear to be influenced by the different lighting regimes, however, a greater relative crystallinity measured by wide-angle X-ray scattering and greater crystalline quality as judged by differential scanning calorimetry was observed under the diurnal lighting regime. NBS cultivated under the diurnal photosynthetic lighting regime displayed lower amylose content (18.7%), and shorter amylose chains than its counterpart grown under constant light. Although the relati...
Plant science : an international journal of experimental plant biology, 2017
Grasses such as wheat and barley constitute some of the main crops currently cultivated worldwide... more Grasses such as wheat and barley constitute some of the main crops currently cultivated worldwide. Cultivation of wild cereals such as Hordeum spontaneum (wild barley) and Triticum diccocoides (wild emmer) started in southwest Asia. Despite the many studies on the origins of agriculture and plant domestication, surprisingly few studies have discussed the importance of the nutritional quality of barley and emmer wheat in their evolution. A comparison of domesticated forms of these cereals with their wild progenitors suggests an evolution towards bigger grains with higher glycemic loads in the form of higher relative starch content with lower relative protein, fiber and mineral contents. In this work we hypothesize that in addition to the simple explanation that larger grains emerged under domestication, complex and indirect effects such as increased glycemic index and sweet taste should also be taken into consideration.
The plant-specific NAC transcription factors have attracted particular attention because of their... more The plant-specific NAC transcription factors have attracted particular attention because of their involvement in stress responses, senescence, and nutrient remobilization. The HvNAC005 gene of barley encodes a protein belonging to subgroup NAC-a6 of the NAC family. This study shows that HvNAC005 is associated with developmental senescence. It was significantly up-regulated following ABA treatment, supported by ABA-responsive elements in its promoter, but it was not up-regulated during dark-induced senescence. The C-termini of proteins closely related to HvNAC005 showed overall high divergence but also contained conserved short motifs. A serine- and leucine-containing central motif was essential for transcriptional activity of the HvNAC005 C-terminus in yeast. Over-expression of HvNAC005 in barley resulted in a strong phenotype with delayed development combined with precocious senescence. The over-expressing plants showed up-regulation of genes involved with secondary metabolism, hor...
Users may download and print one copy of any publication from the public portal for the purpose... more Users may download and print one copy of any publication from the public portal for the purpose of private study or research. You may not further distribute the material or use it for any profit-making activity or commercial gain You may freely distribute the URL identifying the publication in the public portal If you believe that this document breaches copyright please contact us providing details, and we will remove access to the work immediately and investigate your claim.
International journal of biological macromolecules, 2016
The effects of amylose deposition on crystalline regions of barley starch granules were studied i... more The effects of amylose deposition on crystalline regions of barley starch granules were studied in granules containing zero to 99.1% amylose using "waxy" (WBS, 0% amylose), normal (NBS, 18% amylose) and amylose-only barley lines (AOS, 99.1% amylose). The effects were probed after hydrolysis of amorphous regions of starch granules in dilute HCl generating lintners, which typically represent the crystalline lamella of starch granules. Compared to NBS and WBS, AOS granules exhibited an irregular, multilobular morphology with a rough surface texture. AOS displayed lower rates of acid hydrolysis than WBS, and AOS reached a plateau at ∼45wt% acid hydrolysis. High-performance anion-exchange chromatography of lintners at equivalent levels of hydrolysis (45wt%) revealed the average degree of polymerization (DP) of AOS lintners was 21, substantially smaller than that of NBS and WBS (DP 42). AOS lintners contained the lowest number of chains (NC) per molecule (1.1) compared to NBS (2...
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