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Background: Hemp (Cannabis sativa) is a crop with a wide range of uses, from the production of fiber and seeds to the secondary metabolites for medicinal purposes. In addition, it is characterized by high biomass yield and the ability to accumulate heavy metals, which makes this plant convenient for phytoremediation purposes. In this study, the effect of applying exogenous biodegradable chelating agents, citric acid (CA) and nitrilotriacetic acid (NTA), to zinc-contaminated soil on zinc (Zn) uptake by two industrial hemp varieties 'Felina 32' and 'Monoica' was studied. The effect of CA and NTA on available Zn in soils was investigated using an 'in pot' experiment under controlled conditions. The effect of both tested compounds on soil microbial activity was simultaneously evaluated. Results: After the application of NTA at a concentration of 5 mmol LMINUS SIGN 1, a > threefold increased accumulation of Zn in the above-ground parts was recorded in the 'Felina 32' variety. In the 'Monoica' variety, the levels of Zn in the above-ground parts were increased > twofold. NTA affected the soil microbiome negatively, causing decreased enzyme activity (in 'Monoica' planted soil) and induced respiration (in 'Monoica' and especially in 'Felina 32' planted soil). On the other hand, CA application did not lead to significantly increased Zn levels in any of the studied hemp varieties. Together with CA's negative effects on some soil enzymes, CA enhanced urease activity, dehydrogenase and several respiration types for the 'Felina 32' variety and exerted less detrimental effect on the soil microbiome. No toxic effects from increased Zn uptake and accumulation in experimental plants were detected, accounting for the unchanged physiological stress markers (levels of photosynthetic pigments and proline in leaves, chlorophyll fluorescence parameters) and selected growth traits of the above-ground organs and root system. Conclusions: From the studied varieties, 'Felina 32' seems to be mo, info:eu-repo/semantics/openAccess

In Arabidopsis, the plastidial isoform of phosphoglucose isomerase, PGI1, mediates growth and photosynthesis, likely due to its involvement in the vascular production of cytokinins (CK). To examine this hypothesis, we characterized pgi1-2 knockout plants impaired in PGI1 and pgi1-2 plants specifically expressing PGI1 in root tips and vascular tissues. Moreover, to investigate whether the phenotype of pgi1-2 plants is due to impairments in the plastidial oxidative pentose phosphate pathway (OPPP) or the glycolytic pathway, we characterized pgl3-1 plants with reduced OPPP and pfk4pfk5 knockout plants impaired in plastidial glycolysis. Compared with wild-type (WT) leaves, pgi1-2 leaves exhibited weaker expression of photosynthesis- and 2-C-methyl-D-erythritol 4-P (MEP) pathway-related proteins, and stronger expression of oxidative stress protection-related enzymes. Consistently, pgi1-2 leaves accumulated lower levels of chlorophyll, and higher levels of tocopherols, flavonols and anthocyanins than the WT. Vascular- and root tip-specific PGI1 expression countered the reduced photosynthesis, low MEP pathway-derived CK content, dwarf phenotype and the metabolic characteristics of pgi1-2 plants, reverting them to WT-like levels. Moreover, pgl3-1, but not pfk4pfk5 plants phenocopied pgi1-2. Histochemical analyses of plants expressing GUS under the control of promoter regions of genes encoding plastidial OPPP enzymes exhibited strong GUS activity in root tips and vascular tissues. Overall, our findings show that root tip and vascular PGI1-mediated plastidial OPPP activity affects photosynthesis and growth through mechanisms involving long-distance modulation of the leaf proteome by MEP pathway-derived CKs.

Background: Hemp (Cannabis sativa) is a crop with a wide range of uses, from the production of fiber and seeds to the secondary metabolites for medicinal purposes. In addition, it is characterized by high biomass yield and the ability to accumulate heavy metals, which makes this plant convenient for phytoremediation purposes. In this study, the effect of applying exogenous biodegradable chelating agents, citric acid (CA) and nitrilotriacetic acid (NTA), to zinc-contaminated soil on zinc (Zn) uptake by two industrial hemp varieties 'Felina 32' and 'Monoica' was studied. The effect of CA and NTA on available Zn in soils was investigated using an 'in pot' experiment under controlled conditions. The effect of both tested compounds on soil microbial activity was simultaneously evaluated. Results: After the application of NTA at a concentration of 5 mmol LMINUS SIGN 1, a > threefold increased accumulation of Zn in the above-ground parts was recorded in the 'Felina 32' variety. In the 'Monoica' variety, the levels of Zn in the above-ground parts were increased > twofold. NTA affected the soil microbiome negatively, causing decreased enzyme activity (in 'Monoica' planted soil) and induced respiration (in 'Monoica' and especially in 'Felina 32' planted soil). On the other hand, CA application did not lead to significantly increased Zn levels in any of the studied hemp varieties. Together with CA's negative effects on some soil enzymes, CA enhanced urease activity, dehydrogenase and several respiration types for the 'Felina 32' variety and exerted less detrimental effect on the soil microbiome. No toxic effects from increased Zn uptake and accumulation in experimental plants were detected, accounting for the unchanged physiological stress markers (levels of photosynthetic pigments and proline in leaves, chlorophyll fluorescence parameters) and selected growth traits of the above-ground organs and root system. Conclusions: From the studied varieties, 'Felina 32' seems to be mo, info:eu-repo/semantics/openAccess

Background: Hemp (Cannabis sativa) is a crop with a wide range of uses, from the production of fiber and seeds to the secondary metabolites for medicinal purposes. In addition, it is characterized by high biomass yield and the ability to accumulate heavy metals, which makes this plant convenient for phytoremediation purposes. In this study, the effect of applying exogenous biodegradable chelating agents, citric acid (CA) and nitrilotriacetic acid (NTA), to zinc-contaminated soil on zinc (Zn) uptake by two industrial hemp varieties 'Felina 32' and 'Monoica' was studied. The effect of CA and NTA on available Zn in soils was investigated using an 'in pot' experiment under controlled conditions. The effect of both tested compounds on soil microbial activity was simultaneously evaluated. Results: After the application of NTA at a concentration of 5 mmol LMINUS SIGN 1, a > threefold increased accumulation of Zn in the above-ground parts was recorded in the 'Felina 32' variety. In the 'Monoica' variety, the levels of Zn in the above-ground parts were increased > twofold. NTA affected the soil microbiome negatively, causing decreased enzyme activity (in 'Monoica' planted soil) and induced respiration (in 'Monoica' and especially in 'Felina 32' planted soil). On the other hand, CA application did not lead to significantly increased Zn levels in any of the studied hemp varieties. Together with CA's negative effects on some soil enzymes, CA enhanced urease activity, dehydrogenase and several respiration types for the 'Felina 32' variety and exerted less detrimental effect on the soil microbiome. No toxic effects from increased Zn uptake and accumulation in experimental plants were detected, accounting for the unchanged physiological stress markers (levels of photosynthetic pigments and proline in leaves, chlorophyll fluorescence parameters) and selected growth traits of the above-ground organs and root system. Conclusions: From the studied varieties, 'Felina 32' seems to be mo

Background: Hemp (Cannabis sativa) is a crop with a wide range of uses, from the production of fiber and seeds to the secondary metabolites for medicinal purposes. In addition, it is characterized by high biomass yield and the ability to accumulate heavy metals, which makes this plant convenient for phytoremediation purposes. In this study, the effect of applying exogenous biodegradable chelating agents, citric acid (CA) and nitrilotriacetic acid (NTA), to zinc-contaminated soil on zinc (Zn) uptake by two industrial hemp varieties 'Felina 32' and 'Monoica' was studied. The effect of CA and NTA on available Zn in soils was investigated using an 'in pot' experiment under controlled conditions. The effect of both tested compounds on soil microbial activity was simultaneously evaluated. Results: After the application of NTA at a concentration of 5 mmol LMINUS SIGN 1, a > threefold increased accumulation of Zn in the above-ground parts was recorded in the 'Felina 32' variety. In the 'Monoica' variety, the levels of Zn in the above-ground parts were increased > twofold. NTA affected the soil microbiome negatively, causing decreased enzyme activity (in 'Monoica' planted soil) and induced respiration (in 'Monoica' and especially in 'Felina 32' planted soil). On the other hand, CA application did not lead to significantly increased Zn levels in any of the studied hemp varieties. Together with CA's negative effects on some soil enzymes, CA enhanced urease activity, dehydrogenase and several respiration types for the 'Felina 32' variety and exerted less detrimental effect on the soil microbiome. No toxic effects from increased Zn uptake and accumulation in experimental plants were detected, accounting for the unchanged physiological stress markers (levels of photosynthetic pigments and proline in leaves, chlorophyll fluorescence parameters) and selected growth traits of the above-ground organs and root system. Conclusions: From the studied varieties, 'Felina 32' seems to be mo, info:eu-repo/semantics/openAccess

Heat and drought events often occur concurrently as a consequence of climate change and have a severe impact on crop growth and yield. Besides, the accumulative increase in the atmospheric CO2 level is expected to be doubled by the end of this century. It is essential to understand the consequences of climate change combined with the CO2 levels on relevant crops such as wheat. This study evaluated the physiology and metabolite changes and grain yield in heat-sensitive (SF29) and heat-tolerant (LM20) wheat genotypes under individual heat stress or combined with drought applied during anthesis at ambient (aCO(2)) and elevated CO2 (eCO(2)) levels. Both genotypes enhanced similarly the WUE under combined stresses at eCO(2). However, this increase was due to different stress responses, whereas eCO(2) improved the tolerance in heat-sensitive SF29 by enhancing the gas exchange parameters, and the accumulation of compatible solutes included glucose, fructose, beta-alanine, and GABA to keep water balance; the heat-tolerant LM20 improved the accumulation of phosphate and sulfate and reduced the lysine metabolism and other metabolites including N-acetylornithine. These changes did not help the plants to improve the final yield under combined stresses at eCO(2). Under non-stress conditions, eCO(2) improved the yield of both genotypes. However, the response differed among genotypes, most probably as a consequence of the eCO(2)-induced changes in glucose and fructose at anthesis. Whereas the less-productive genotype LM20 reduced the glucose and fructose and increased the grain dimension as the effect of the eCO(2) application, the most productive genotype SF29 increased the two carbohydrate contents and ended with higher weight in the spikes. Altogether, these findings showed that the eCO(2) improves the tolerance to combined heat and drought stress but not the yield in spring wheat under stress conditions through different mechanisms. However, under non-stress conditions, it

Heat and drought events often occur concurrently as a consequence of climate change and have a severe impact on crop growth and yield. Besides, the accumulative increase in the atmospheric CO2 level is expected to be doubled by the end of this century. It is essential to understand the consequences of climate change combined with the CO2 levels on relevant crops such as wheat. This study evaluated the physiology and metabolite changes and grain yield in heat-sensitive (SF29) and heat-tolerant (LM20) wheat genotypes under individual heat stress or combined with drought applied during anthesis at ambient (aCO(2)) and elevated CO2 (eCO(2)) levels. Both genotypes enhanced similarly the WUE under combined stresses at eCO(2). However, this increase was due to different stress responses, whereas eCO(2) improved the tolerance in heat-sensitive SF29 by enhancing the gas exchange parameters, and the accumulation of compatible solutes included glucose, fructose, beta-alanine, and GABA to keep water balance; the heat-tolerant LM20 improved the accumulation of phosphate and sulfate and reduced the lysine metabolism and other metabolites including N-acetylornithine. These changes did not help the plants to improve the final yield under combined stresses at eCO(2). Under non-stress conditions, eCO(2) improved the yield of both genotypes. However, the response differed among genotypes, most probably as a consequence of the eCO(2)-induced changes in glucose and fructose at anthesis. Whereas the less-productive genotype LM20 reduced the glucose and fructose and increased the grain dimension as the effect of the eCO(2) application, the most productive genotype SF29 increased the two carbohydrate contents and ended with higher weight in the spikes. Altogether, these findings showed that the eCO(2) improves the tolerance to combined heat and drought stress but not the yield in spring wheat under stress conditions through different mechanisms. However, under non-stress conditions, it

Item does not contain fulltext, BACKGROUND Strigolactones (SLs) are plant hormones that play various roles in plant development. The chemical stability of SLs depends on the solvent, the pH, and the presence of nucleophiles. Hydrolysis leads to detachment of the butenolide ring, and plays a crucial role in the initial stages of the signal-transduction process occurring between the receptor and the SL signaling molecule. RESULTS To date, two different mechanisms have been proposed for SL hydrolysis. Results obtained from kinetic, thermodynamic, and mass spectral data for the reaction between the widely used synthetic SL analog GR24 and seven different nucleophiles demonstrate that the reaction proceeds via the Michael addition?elimination mechanism. CONCLUSION This study provides valuable information on the chemical stability of GR24 in different plant growth media and buffers. Such information is valuable for scientists using GR24 treatments to study SL-regulated processes in plants. ? 2017 Society of Chemical Industry

Item does not contain fulltext, BACKGROUND Strigolactones (SLs) are plant hormones that play various roles in plant development. The chemical stability of SLs depends on the solvent, the pH, and the presence of nucleophiles. Hydrolysis leads to detachment of the butenolide ring, and plays a crucial role in the initial stages of the signal-transduction process occurring between the receptor and the SL signaling molecule. RESULTS To date, two different mechanisms have been proposed for SL hydrolysis. Results obtained from kinetic, thermodynamic, and mass spectral data for the reaction between the widely used synthetic SL analog GR24 and seven different nucleophiles demonstrate that the reaction proceeds via the Michael addition?elimination mechanism. CONCLUSION This study provides valuable information on the chemical stability of GR24 in different plant growth media and buffers. Such information is valuable for scientists using GR24 treatments to study SL-regulated processes in plants. ? 2017 Society of Chemical Industry

LONELY GUY (LOG) genes encode cytokinin riboside 5′-monophosphate phosphoribohydrolases and are directly involved in the activation of cytokinins. To assess whether LOG proteins affect the influence of cytokinin on nodulation, we studied two LOG genes of Medicago truncatula. Expression analysis showed that MtLOG1 and MtLOG2 were upregulated during nodulation in a CRE1-dependent manner. Expression was mainly localized in the dividing cells of the nodule primordium. In addition, RNA interference revealed that MtLOG1 is involved in nodule development and that the gene plays a negative role in lateral root development. Ectopic expression of MtLOG1 resulted in a change in cytokinin homeostasis, triggered cytokinin-inducible genes and produced roots with enlarged vascular tissues and shortened primary roots. In addition, those 35S:LOG1 roots also displayed fewer nodules than the wild-type. This inhibition in nodule formation was local, independent of the SUPER NUMERIC NODULES gene, but coincided with an upregulation of the MtCLE13 gene, encoding a CLAVATA3/EMBRYO SURROUNDING REGION peptide. In conclusion, we demonstrate that in M. truncatula LOG proteins might be implicated in nodule primordium development and lateral root formation.

Together, auxin and cytokinin regulate many of the processes that are critical to plant growth, development, and environmental responsiveness. We have previously shown that exogenous auxin regulates cytokinin biosynthesis in Arabidopsis thaliana. In this work, we show that, conversely, the application or induced ectopic biosynthesis of cytokinin leads to a rapid increase in auxin biosynthesis in young, developing root and shoot tissues. We also show that reducing endogenous cytokinin levels, either through the induction of CYTOKININ OXIDASE expression or the mutation of one or more of the cytokinin biosynthetic ISOPENTENYLTRANSFERASE genes leads to a reduction in auxin biosynthesis. Cytokinin modifies the abundance of transcripts for several putative auxin biosynthetic genes, suggesting a direct induction of auxin biosynthesis by cytokinin. Our data indicate that cytokinin is essential, not only to maintain basal levels of auxin biosynthesis in developing root and shoot tissues but also for the dynamic regulation of auxin biosynthesis in response to changing developmental or environmental conditions. In combination with our previous work, the data suggest that a homeostatic feedback regulatory loop involving both auxin and cytokinin signaling acts to maintain appropriate auxin and cytokinin concentrations in developing root and shoot tissues.

An improved method for determining the relative biosynthetic rate of isoprenoid cytokinins has been developed. A set of 11 relevant isoprenoid cytokinins, including zeatin isomers, was separated by ultra performance liquid chromatography in less than 6 min. The iP-type cytokinins were observed to give rise to a previously-unknown fragment at m/z 69; we suggest that the diagnostic (204-69) transition can be used to monitor the biosynthetic rate of isopentenyladenine. Furthermore, we found that by treating the cytokinin nucleotides with alkaline phosphatase prior to analysis, the sensitivity of the detection process could be increased. In addition, derivatization (propionylation) improved the ESI-MS response by increasing the analytes' hydrophobicity. Indeed, the ESI-MS response of propionylated isopentenyladenosine was about 34% higher than that of its underivatized counterpart. Moreover, the response of the derivatized zeatin ribosides was about 75% higher than that of underivatized zeatin ribosides. Finally, we created a web-based calculator (IZOTOP) that facilitates MS/MS data processing and offer it freely to the research community.

We have developed a method for analyzing polar compounds by reversed-phase LC-ESI-MS following esterification of the analytes' free hydroxyl groups with propionyl or benzoyl acid anhydride. The method was applied to members of the plant hormone group cytokinins, which includes adenine bases, ribosides/glycosides, and nucleotides substituted at N-6 with an isoprenoid side chain, spanning a wide range of polarity. It was also used to analyze other compounds of biological importance, e.g., the nucleotides AMP, ADP, and ATP. The formation of more hydrophobic derivatives had a significant impact on two aspects of the analysis. The retention on a reversed-phase material was greatly increased without the use of any acetate/formate buffer or ion pairing reagent, and the ESI response was enhanced, due to the higher surface activities of the derivatives. Detection limits of propionylated cytokinins were in the high-attomole to low-femtomole range, an improvement by factors of 10-100 compared to previously reported figures. Using an automated SPE-based purification method, 12 endogenous cytokinins were quantified in extracts from 20- to 100-mg samples of leaves (from the plant Arabidopsis thaliana) with high accuracy and precision. Furthermore, the chromatographic properties of the benzoylated AMP, ADP, and ATP in the reversed-phase LC-MS system were much better in terms of retention, separation, and sensitivity than those of their underivatized counterparts, even without the use of any ion pairing reagent. Our data show that derivatization followed by LC-ESI-MS is an effective strategy for analyzing low molecular weight compounds, enabling compounds with a wide range of polarity to be determined in a single-injection LC-MS analysis.

One of the most long-lived models in plant science is the belief that the long-distance transport and ratio of two plant hormones, auxin and cytokinin, at the site of action control major developmental events such as apical dominance. We have used in vivo deuterium labeling and mass spectrometry to investigate the dynamics of homeostatic cross talk between the two plant hormones. Interestingly, auxin mediates a very rapid negative control of the cytokinin pool by mainly suppressing the biosynthesis via the isopentenyladenosine-5'-monophosphate-independent pathway. In contrast, the effect of cytokinin overproduction on the entire auxin pool in the plant was slower, indicating that this most likely is mediated through altered development. In addition, we were able to confirm that the lateral root meristems are likely to be the main sites of isopentenyladenosine-5'-monophosphate-dependent cytokinin synthesis, and that the aerial tissue of the plant surprisingly also was a significant source of cytokinin biosynthesis. Our demonstration of shoot-localized synthesis, together with data demonstrating that auxin imposes a very rapid regulation of cytokinin biosynthesis, illustrates that the two hormones can interact also on the metabolic level in controlling plant development, and that the aerial part of the plant has the capacity to synthesize its own cytokinin independent of long-range transport from the root system.