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Salt stress affects plant growth and productivity. In this study we determined the roles of eight genes involved in photosynthesis, using gene co-expression network analysis, under salt-stress conditions using Arabidopsis knockout mutants. The green area of the leaves was minimum in the

Increasing crop productivity is our major challenge if we are to meet global needs for food, fodder and fuel. Controlling the content of the plant hormone cytokinin is a method of improving plant productivity. Cytokinin oxidase/dehydrogenase (CKO/CKX) is a major target in this regard because it degrades cytokinins. Here, we describe the synthesis and biological activities of new CKX inhibitors derived mainly from diphenylurea. They were tested on four CKX isoforms from maize and Arabidopsis, where the best compounds showed IC50 values in the 10–8 M concentration range. The binding mode of the most efficient inhibitors was characterized from high-resolution crystal complexed structures. Although these compounds do not possess intrinsic cytokinin activity, we have demonstrated their tremendous potential for use in the plant tissue culture industry as well as in agriculture. We have identified a key substance, compound 19, which not only increases stress resistance and seed yield in Arabidopsis, but also improves the yield of wheat, barley and rapeseed grains under field conditions. Our findings reveal that modulation of cytokinin levels via CKX inhibition can positively affect plant growth, development and yield, and prove that CKX inhibitors can be an attractive target in plant biotechnology and agriculture.

In Arabidopsis (Arabidopsis thaliana), the plastidial isoform of phosphoglucose isomerase (PGI1) mediates photosynthesis, metabolism, and development, probably due to its involvement in the synthesis of isoprenoid-derived signals in vascular tissues. Microbial volatile compounds (VCs) with molecular masses of, This work was supported by the Ministerio de Ciencia e Innovación (MCIN) and Agencia Estatal de Investigación (AEI)/10.13039/501100011033/ (grants BIO2016-78747-P, PID2019-104685GB-100 and PID2019-107657RB-C22) and the Ministry of Education, Youth and Sport of the Czech Republic and the European Regional Development Fund (ERDF) project entitled “Plants as a tool for sustainable global development” (No. CZ.02.1.01/0.0/0.0/16_019/0000827).

Aims The aim of this work was evaluating the interaction between two of the most widely used strains for soybean inoculation in Argentina, B. japonicum E109 (BjE109) and A. brasilense Az39 (AbAz39). Methods Five treatments were performed: (i) uninoculated seeds; (ii) seeds inoculated with BjE109; (iii) seeds inoculated with AbAz39; (iv) seeds co-inoculated with BjE109 and AbAz39 in a 1:1 proportion (BjE109 + AbAz39) at the seeds sowing and (v) seeds inoculated with a 1:1 proportion of BjE109 and AbAz39 (BjE109-AbAz39) 24 h before seeds sowing. Each treatment was assessed through a seed recovery assay, glasshouse assays and field assays. Results The combination between the two strains improved the ability of BjE109 to survive on soybean seeds after inoculation partially explained by AbAz39’s capacity to produce diverse biologically active molecules. As a result of the greater rhizobial survival on seeds the nodulation values and symbiosis parameters like nodule number, size and biomass and nodulation percentage also increased. In agreement with these observations, combining BjE109 and AbAz39 at strains the grain yield under field conditions were 13,3 and 17,3% greater than single BjE109 inoculation. Conclusions These results here show that the pre-culture combining BjE109 and AbAz39 before the inoculation to the soybean seeds has benefits in plant nodulation and hence production, more than individual inoculation with BjE109 or AbAz39, or the immediate co-inoculation of both strains.

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 CO

Commercial interest in biostimulants as a tool for sustainable green economics and agriculture concepts is on a steep rise, being followed by increasing demand to employ efficient scientific methods to develop new products and understand their mechanisms of action. Biostimulants represent a highly diverse group of agents derived from various natural sources. Regardless of their nutrition content and composition, they are classified by their ability to improve crop performance through enhanced nutrient use efficiency, abiotic stress tolerance, and quality of crops. Numerous reports have described modern, non-invasive sensor-based phenotyping methods in plant research. This review focuses on applying phenotyping approaches in biostimulant research and development, and maps the evolution of interaction of these two intensively growing domains. How phenotyping served to identify new biostimulants, the description of their biological activity, and the mechanism/mode of action are summarized. Special attention is dedicated to the indoor high-throughput methods using model plants suitable for biostimulant screening and developmental pipelines, and high-precision approaches used to determine biostimulant activity. The need for a complex method of testing biostimulants as multicomponent products through integrating other -omic approaches followed by advanced statistical/mathematical tools is emphasized.

Plant phenomics is becoming a common tool employed to characterize the mode of action of biostimulants. A combination of this technique with other omics such as metabolomics can offer a deeper understanding of a biostimulant effect

Mentha species are widely used as food, medicine, spices, and flavoring agents. Thus, chemical composition is an important parameter for assessing the quality of mints. In general, the contents of menthol, menthone, eucalyptol, and limonene comprise one of the major parameters for assessing the quality of commercially important mints. Building further on the phytochemical characterization of the quality of Mentha species, this work was focused on the composition of phenolic compounds in methanolic extracts. Thirteen Mentha species were grown under the same environmental conditions, and their methanolic extracts were subjected to the LC–MS/MS (liquid chromatography–tandem mass spectrometry) profiling of phenolics and the testing their biological activities, i.e., antioxidant and tyrosinase inhibition activities, which are important features for the cosmetic industry. The total phenolic content (TPC) ranged from 14.81 ± 1.09 mg GAE (gallic acid equivalents)/g for Mentha cervina to 58.93. ± 8.39 mg GAE/g for Mentha suaveolens. The antioxidant activity of examined Mentha related with the content of the phenolic compounds and ranged from 22.79 ± 1.85 to 106.04 ± 3.26 mg TE (Trolox equivalents)/g for M. cervina and Mentha x villosa, respectively. Additionally, Mentha pulegium (123.89 ± 5.64 mg KAE (kojic acid equivalents)/g) and Mentha x piperita (102.82 ± 15.16 mg KAE/g) showed a strong inhibition of the enzyme tyrosinase, which is related to skin hyperpigmentation. The most abundant compound in all samples was rosmarinic acid, ranging from 1363.38 ± 8323 to 2557.08 ± 64.21 μg/g. In general, the levels of phenolic acids in all examined mint extracts did not significantly differ. On the contrary, the levels of flavonoids varied within the species, especially in the case of hesperidin (from 0.73 ± 0.02 to 109. 39 ± 2.01 μg/g), luteolin (from 1.84 ± 0.11 to 31.03 ± 0.16 μg/g), and kaempferol (from 1.30 ± 0.17 to 33.68 ± 0.81 μg/g). Overall results indicated that all examined mints possess significant amounts of phenolic compounds that are responsible for antioxidant activity and, to some extent, for tyrosinase inhibition activity. Phenolics also proved to be adequate compounds, together with terpenoids, for the characterization of Mentha sp. Additionally, citrus-scented Mentha x villosa could be selected as a good candidate for the food and pharmaceutical industry, especially due its chemical composition and easy cultivation, even in winter continental conditions.

Summary Drought stress impacts the quality and yield of Pisum sativum. Here, we show how short periods of limited water availability during the vegetative stage of pea alters phloem sap content and how these changes are connected to strategies used by plants to cope with water deficit. We have investigated the metabolic content of phloem sap exudates and explored how this reflects P. sativum physiological and developmental responses to drought. Our data show that drought is accompanied by phloem‐mediated redirection of the components that are necessary for cellular respiration and the proper maintenance of carbon/nitrogen balance during stress. The metabolic content of phloem sap reveals a shift from anabolic to catabolic processes as well as the developmental plasticity of P. sativum plants subjected to drought. Our study underlines the importance of phloem‐mediated transport for plant adaptation to unfavourable environmental conditions. We also show that phloem exudate analysis can be used as a useful proxy to study stress responses in plants. We propose that the decrease in oleic acid content within phloem sap could be considered as a potential marker of early signalling events mediating drought response., Significance Statement We describe the changes in phloem metabolic content of pea (Pisum sativum) plants subjected to drought and show how the observed patterns reflect physiological and developmental responses. Phloem exudate analysis itself gives us a clear and informative picture of the stress response strategy centred on above‐ground growth reduction and the reuse and redistribution of carbon and nitrogen. This provides evidence for the importance of phloem‐mediated long‐distance coordination in plant responses to drought. Fluctuations of particular metabolites in vascular sap can be used to monitor drought response in pea plants.

Nanomaterials-mediated contamination (including the highly reactive metal oxides ZnO nanoparticles) is becoming one of the most concerning issues worldwide. In this study, the toxic effects of two chemical species of Zn (ZnO nanoparticles and bulk ZnSO

This work presents a pipeline for optimizing semi-hydroponic growth conditions and analyzes the impact on the growth and metabolism in three Mentha species (M. arvensis, M. x piperita, or M. spicata) and three Ocimum basilicum genotypes (́Chládek červená́, ́Litrá, or ́Máneś). The plant growth and the content- of nitrogen-containing compounds, phenolics, and terpenoids were determined under different nitrate concentrations and salt stress. Different responses were observed among genotypes for both species. ́Chládek červená́ had the best growth under low nitrate, with lower histamine and higher flavonoid levels. Mentha x piperita was the best mint species performing under low nitrate and salt stress. Altogether, we demonstrate that a combination of phenomics and metabolomics is ideal to identify the optimal growth conditions for these plants and the chemical markers associated with these conditions. Besides, we showed that both primary and secondary metabolites can be good markers for classifying both species.

Drought stress limits plant growth and productivity. It triggers many responses by inducing changes in plant morphology and physiology. KDML105 rice is a key rice variety in Thailand and is normally grown in the northeastern part of the country. The chromosome segment substitution lines (CSSLs) were developed by transferring putative drought tolerance loci (QTLs) on chromosome 1, 3, 4, 8, or 9 into the KDML105 rice genome. CSSL104 is a drought-tolerant line with higher net photosynthesis and leaf water potential than KDML105 rice. The analysis of CSSL104 gene regulation identified the loci associated with these traits via gene co-expression network analysis. Most of the predicted genes are involved in the photosynthesis process. These genes are also conserved in Arabidopsis thaliana. Seven genes encoding chloroplast proteins were selected for further analysis through characterization of Arabidopsis tagged mutants. The response of these mutants to drought stress was analyzed daily for seven days after treatment by scoring green tissue areas via the PlantScreen&trade, XYZ system. Mutation of these genes affected green areas of the plant and stability index under drought stress, suggesting their involvement in drought tolerance.

Volatile compounds (VCs) emitted by the fungal phytopathogen Penicillium aurantiogriseum promote root growth and developmental changes in Arabidopsis. Here we characterized the metabolic and molecular responses of roots to fungal volatiles. Proteomic analyses revealed that these compounds reduce the levels of aquaporins, the iron carrier IRT1 and apoplastic peroxidases. Fungal VCs also increased the levels of enzymes involved in the production of mevalonate (MVA)‐derived isoprenoids, nitrogen assimilation and conversion of methionine to ethylene and cyanide. Consistently, fungal VC‐treated roots accumulated high levels of hydrogen peroxide (H2O2), MVA‐derived cytokinins, ethylene, cyanide and long‐distance nitrogen transport amino acids. qRT‐PCR analyses showed that many proteins differentially expressed by fungal VCs are encoded by VC non‐responsive genes. Expression patterns of hormone reporters and developmental characterization of mutants provided evidence for the involvement of cyanide scavenging and enhanced auxin, ethylene, cytokinin and H2O2 signaling in the root architecture changes promoted by fungal VCs. Our findings show that VCs from P. aurantiogriseum modify root metabolism and architecture, and improve nutrient and water use efficiencies through transcriptionally and non‐transcriptionally regulated proteome resetting mechanisms. Some of these mechanisms are subject to long‐distance regulation by photosynthesis and differ from those triggered by VCs emitted by beneficial microorganisms.

Abscisic acid (ABA), auxins, and cytokinins (CKs) are known to be closely linked to nitrogen signaling. In particular, CKs control the effects of nitrate availability on plant growth. Our group has shown that treatment with high nitrate concentrations limits root growth and leaf development in maize, and conditions the development of younger roots and leaves. CKs also affect source-sink relationships in plants. Based on these results, we hypothesized that CKs regulate the source-sink relationship in maize via a mechanism involving complex crosstalk with the main auxin indole-3-acetic acid (IAA) and ABA. To evaluate this hypothesis, various CK metabolites, IAA, and ABA were quantified in the roots and in source and sink leaves of maize plants treated with high and normal nitrate concentrations. The data obtained suggest that the cis and trans isomers of zeatin play completely distinct roles in maize growth regulation by a complex crosstalk with IAA and ABA. We demonstrate that while trans-zeatin (tZ) and isopentenyladenine (iP) regulate nitrate uptake and thus control final leaf sizes, cis-zeatin (cZ) regulates source and sink strength, and thus controls leaf development. The implications of these findings relating to the roles of ABA and IAA in plants’ responses to varying nitrate concentrations are also discussed.

Nanomaterials-mediated contamination (including the highly reactive metal oxides ZnO nanoparticles) is becoming one of the most concerning issues worldwide. In this study, the toxic effects of two chemical species of Zn (ZnO nanoparticles and bulk ZnSO4) were investigated in bean plants, following either foliar or soil application, at concentrations from 250 to 2000 mg L−1 using biochemical assays, proteomics and metabolomics. The accumulation of Zn in plant tissues depended on the application type, zinc chemical form and concentration, in turn triggering distinctive morphological, physiological, and redox responses. Bean plants were more sensitive to the foliar than to the soil application, and high concentrations of ZnO NP and bulk ZnSO4 determined the highest plant growth inhibition and stress symptoms. However, low dosages of ZnSO4 induced a slight plant growth promotion and better physiological and antioxidative response. Low concentration of Zn leaded to increased activity of stress-related proteins and secondary metabolites with antioxidant capacity, while increasing concentration reached the exhausted phase of the plant stress response, reducing the antioxidant defense system. Such high concentrations increased lipids peroxidation, protein degradation and membranes integrity. Oxidative damage occurred at high concentrations of both chemical species of Zn. Foliar spraying impaired photosynthetic efficiency, while soil applications (especially ZnSO4) elicited antioxidant metabolites and proteins, and impaired chloroplast-related proteins involved in the electron transport chain and ATP production. Taken together, the results highlighted distinctive and nanoparticles-related toxic effects of ZnO in bean, compared to ionic forms of Zn.

Microorganisms produce volatile compounds (VCs) that promote plant growth and photosynthesis through complex mechanisms involving cytokinin (CK) and abscisic acid (ABA). We hypothesized that plants' responses to microbial VCs involve posttranslational modifications of the thiol redox proteome through action of plastidial NADPH‐dependent thioredoxin reductase C (NTRC), which regulates chloroplast redox status via its functional relationship with 2‐Cys peroxiredoxins. To test this hypothesis, we analysed developmental, metabolic, hormonal, genetic, and redox proteomic responses of wild‐type (WT) plants and a NTRC knockout mutant (ntrc) to VCs emitted by the phytopathogen Alternaria alternata. Fungal VC‐promoted growth, changes in root architecture, shifts in expression of VC‐responsive CK‐ and ABA‐regulated genes, and increases in photosynthetic capacity were substantially weaker in ntrc plants than in WT plants. As in WT plants, fungal VCs strongly promoted growth, chlorophyll accumulation, and photosynthesis in ntrc–Δ2cp plants with reduced 2‐Cys peroxiredoxin expression. OxiTRAQ‐based quantitative and site‐specific redox proteomic analyses revealed that VCs promote global reduction of the thiol redox proteome (especially of photosynthesis‐related proteins) of WT leaves but its oxidation in ntrc leaves. Our findings show that NTRC is an important mediator of plant responses to microbial VCs through mechanisms involving global thiol redox proteome changes that affect photosynthesis., This work was supported by the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain; Grants BIO2013‐49125‐C2‐1‐P, BIO2016‐78747‐P, and BIO2017‐85195‐C2‐1‐P), the Government of Navarra (Refs. P1044 AGROESTI and P1004 PROMEBIO), the KAKENHI Grants‐in‐Aid for Scientific Research (A; 15H02486) from the Japan Society for the Promotion of Sciences, Strategic International Collaborative Research Program by the Japan Science and Technology Agency (JST SICORP), the Grant for Promotion of KAAB Projects (Niigata University) from the Ministry of Education, Culture, Sports, Science and Technology (Japan) and the Ministry of Education, Youth and Sport of the Czech Republic, and European Regional Development Fund project “Plants as a tool for sustainable global development” (CZ.02.1.01/0.0/0.0/16_019/0000827).

It is known that volatile emissions from some beneficial rhizosphere microorganisms promote plant growth. Here we show that volatile compounds (VCs) emitted by phylogenetically diverse rhizosphere and non-rhizhosphere bacteria and fungi (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote growth and flowering of various plant species, including crops. In Arabidopsis plants exposed to VCs emitted by the phytopathogen Alternaria alternata, changes included enhancement of photosynthesis and accumulation of high levels of cytokinins (CKs) and sugars. Evidence obtained using transgenic Arabidopsis plants with altered CK status show that CKs play essential roles in this phenomenon, because growth and flowering responses to the VCs were reduced in mutants with CK-deficiency (35S:AtCKX1) or low receptor sensitivity (ahk2/3). Further, we demonstrate that the plant responses to fungal VCs are light-dependent. Transcriptomic analyses of Arabidopsis leaves exposed to A. alternata VCs revealed changes in the expression of light- and CK-responsive genes involved in photosynthesis, growth and flowering. Notably, many genes differentially expressed in plants treated with fungal VCs were also differentially expressed in plants exposed to VCs emitted by the plant growth promoting rhizobacterium Bacillus subtilis GB03, suggesting that plants react to microbial VCs through highly conserved regulatory mechanisms.

Nitrogen is one of the main factors that affect plant growth and development. However, high nitrogen concentrations can inhibit both shoot and root growth, even though the processes involved in this inhibition are still unknown. The aim of this work was to identify the metabolic alterations that induce the inhibition of root growth caused by high nitrate supply, when the whole plant growth is also reduced. High nitrate altered nitrogen and carbon metabolism, reducing the content of sugars and inducing the accumulation of Ca2+ and amino acids, such as glutamate, alanine and γ-aminobutyrate (GABA), that could act to replenish the succinate pool in the tricarboxylic acid cycle and maintain its activity. Other metabolic alterations found were the accumulation of the polyamines spermidine and spermine, and the reduction of jasmonic acid (JA) and the ethylene precursor aminocyclopropane-1-carboxylic acid (ACC). These results indicate that the growth root inhibition by high NO3- is a complex metabolic response that involves GABA as a key link between C and N metabolism which, together with plant growth regulators such as auxins, cytokinins, abscisic acid, JA, and the ethylene precursor ACC, is able to regulate the metabolic response of root grown under high nitrate concentrations.

The plastid-localized phosphoglucose isomerase isoform PGI1 is an important determinant of growth in Arabidopsis thaliana, likely due to its involvement in the biosynthesis of plastidial isoprenoid-derived hormones. Here, we investigated whether PGI1 also influences seed yields. PGI1 is strongly expressed in maturing seed embryos and vascular tissues. PGI1-null pgi1-2 plants had ∼60% lower seed yields than wild-type plants, with reduced numbers of inflorescences and thus fewer siliques and seeds per plant. These traits were associated with low bioactive gibberellin (GA) contents. Accordingly, wild-type phenotypes were restored by exogenous GA application. pgi1-2 seeds were lighter and accumulated ∼50% less fatty acids (FAs) and ∼35% less protein than wild-type seeds. Seeds of cytokinin-deficient plants overexpressing CYTOKININ OXIDASE/DEHYDROGENASE1 (35S:AtCKX1) and GA-deficient ga20ox1 ga20ox2 mutants did not accumulate low levels of FAs, and exogenous application of the cytokinin 6-benzylaminopurine and GAs did not rescue the reduced weight and FA content of pgi1-2 seeds. Seeds from reciprocal crosses between pgi1-2 and wild-type plants accumulated wild-type levels of FAs and proteins. Therefore, PGI1 is an important determinant of Arabidopsis seed yield due to its involvement in two processes: GA-mediated reproductive development and the metabolic conversion of plastidial glucose-6-phosphate to storage reserves in the embryo., This work was partially supported by the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain) (Grants BIO2013-49125-C2-1-P and BIO2016-78747-P), the Ministry of Education, Youth, and Sports of the Czech Republic (Grant LO1204 from the National Program of Sustainability I), the Government of Navarra (ref. P1004 PROMEBIO), the Università degli Studi di Milano (UNIMI-RTD-A, Linea2-DBS 2017-2018), and the H2020-MSCA-RISE Project (ExpoSeed GA-691109).

Nitrogen (N) is an essential macronutrient that limits agricultural productivity, and both low and high N supply have been suggested to alter plant growth. The overall aim of this work is to study the impact of nitrate (NO3(-)) in maize yield and the possible causes that induce this alteration. High NO3(-) doses did not increase the yield of maize grown neither in the field nor under controlled conditions. In fact, plants grown under controlled conditions for 45 days with NO3(-) concentrations over 5mM showed a decrease in biomass production. This reduction was perceptible in shoots prior to roots, where phytomer expansion was reduced. Cell size and number were also reduced in the leaves of plants with high NO3(-). This alteration was correlated with the increase of 1-aminocyclopropane-1-carboxylic acid in leaves, which was probably translocated from the roots in order to synthesize ethylene. Cytokinins (CKs) also showed a relevant role in this inhibitory effect, increasing in high NO3(-) plants with a reduction in root and shoot growth, inhibition of apical dominance and a strong decrease of leaf expansion, symptoms described previously as "CK syndrome". We propose that high NO3(-) inhibits maize growth by causing hormonal alterations that modify plant growth from cell to whole plant.

Gamma tocopherol (gT) exhibits beneficial cardiovascular effects partly due to its anti-inflammatory activity. Important sources of gT are vegetable oils. However, little is known to what extent gT can be transferred into marine animal species such as Atlantic salmon by feeding. Therefore, in this study we have investigated the transfer of dietary gT into salmon. To this end, fish were fed a diet supplemented with 170 ppm gT for 16 weeks whereby alpha tocopherol levels were adjusted to 190 ppm in this and the control diet. Feeding gT-rich diets resulted in a three-fold increase in gT concentrations in the liver and fillet compared to non-gT-supplemented controls. Tissue alpha tocopherol levels were not decreased indicating no antagonistic interaction between gamma- and alpha tocopherol in salmon. The concentration of total omega 3 fatty acids slightly increased in response to dietary gT. Furthermore, dietary gT significantly decreased malondialdehyde in the fillet, determined as a biomarker of lipid peroxidation. In the liver of gT fed salmon we observed an overall down-regulation of genes involved in lipid homeostasis. Additionally, gT improved the antioxidant capacity by up-regulating Gpx4a gene expression in the pyloric caeca. We suggest that Atlantic salmon may provide a marine functional source capable of enriching gT for human consumption.

In plants, γ-aminobutyric acid (GABA) accumulates rapidly in response to environmental stress and variations in its endogenous concentration have been shown to affect plant growth. Exogenous application of GABA has also conferred higher stress tolerance by modulating the expression of genes involved in plant signalling, transcriptional regulation, hormone biosynthesis, reactive oxygen species production and polyamine metabolism. Plant hormones play critical roles in adaptation of plants to adverse environmental conditions through a sophisticated crosstalk among them. Several studies have provided evidence for the relationships between GABA, polyamines and hormones such as abscisic acid, cytokinins, auxins, gibberellins and ethylene, among others, focussing on the effect that one specific group of compounds exerts over the metabolic and signalling pathways of others. In this review, we bring together information obtained from plants exposed to several stress conditions and discuss the possible links among these different groups of molecules. The analysis supports the view that highly conserved pathways connect primary and secondary metabolism, with an overlap of regulatory functions related to stress responses and tolerance among phytohormones, amino acids and polyamines.

Nitrogen (N) is an important regulator of photosynthetic carbon (C) flow in plants, and an adequate balance between N and C metabolism is needed for correct plant development. However, an excessive N supply can alter this balance and cause changes in specific organic compounds associated with primary and secondary metabolism, including plant growth regulators. In previous work, we observed that high nitrate supply (15 mM) to maize plants led to a decrease in leaf expansion and overall biomass production, when compared with low nitrate supply (5 mM). Thus, the aim of this work is to study how overdoses of nitrate can affect photosynthesis and plant development. The results show that high nitrate doses greatly increased amino acid production, which led to a decrease in the concentration of 2-oxoglutarate, the main source of C skeletons for N assimilation. The concentration of 1-aminocyclopropane-1-carboxylic acid (and possibly its product, ethylene) also rose in high nitrate plants, leading to a decrease in leaf expansion, reducing the demand for photoassimilates by the growing tissues and causing the accumulation of sugars in source leaves. This accumulation of sugars, together with the decrease in 2-oxoglutarate levels and the reduction in chlorophyll concentration, decreased plant photosynthetic rates. This work provides new insights into how high nitrate concentration alters the balance between C and N metabolism, reducing photosynthetic rates and disrupting whole plant development. These findings are particularly relevant since negative effects of nitrate in contexts other than root growth have rarely been studied.

Sánchez-López, Ángela María et al., Volatile compounds (VCs) emitted by phylogenetically diverse microorganisms (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote photosynthesis, growth, and the accumulation of high levels of starch in leaves through cytokinin (CK)-regulated processes. In Arabidopsis (Arabidopsis thaliana) plants not exposed to VCs, plastidic phosphoglucose isomerase (pPGI) acts as an important determinant of photosynthesis and growth, likely as a consequence of its involvement in the synthesis of plastidic CKs in roots. Moreover, this enzyme plays an important role in connecting the Calvin-Benson cycle with the starch biosynthetic pathway in leaves. To elucidate the mechanisms involved in the responses of plants to microbial VCs and to investigate the extent of pPGI involvement, we characterized pPGI-null pgi1-2 Arabidopsis plants cultured in the presence or absence of VCs emitted by Alternaria alternata. We found that volatile emissions from this fungal phytopathogen promote growth, photosynthesis, and the accumulation of plastidic CKs in pgi1-2 leaves. Notably, the mesophyll cells of pgi1-2 leaves accumulated exceptionally high levels of starch following VC exposure. Proteomic analyses revealed that VCs promote global changes in the expression of proteins involved in photosynthesis, starch metabolism, and growth that can account for the observed responses in pgi1-2 plants. The overall data show that Arabidopsis plants can respond to VCs emitted by phytopathogenic microorganisms by triggering pPGI-independent mechanisms., This work was supported by the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional, Spain (grant nos. BIO2010–18239 and BIO2013–49125–C2–1–P), by the Government of Navarra (grant no. IIM010491.RI1), by the I-Link0939 project from the Ministerio de Economía y Competitividad, by the Ministry of Education, Youth, and Sports of the Czech Republic (grant no. LO1204 from the National Program of Sustainability), by Palacky University institutional support, by predoctoral fellowships from the Spanish Ministry of Science and Innovation (to A.M.S.-L. and P.G.-G.), and by postdoctoral fellowships from the Public University of Navarra (to M.B. and G.A.).

It is known that volatile emissions from some beneficial rhizosphere microorganisms promote plant growth. Here we show that volatile compounds (VCs) emitted by phylogenetically diverse rhizosphere and non-rhizhosphere bacteria and fungi (including plant pathogens and microbes that do not normally interact mutualistically with plants) promote growth and flowering of various plant species, including crops. In Arabidopsis plants exposed to VCs emitted by the phytopathogen Alternaria alternata, changes included enhancement of photosynthesis and accumulation of high levels of cytokinins (CKs) and sugars. Evidence obtained using transgenic Arabidopsis plants with altered CK status show that CKs play essential roles in this phenomenon, because growth and flowering responses to the VCs were reduced in mutants with CK-deficiency (35S:AtCKX1) or low receptor sensitivity (ahk2/3). Further, we demonstrate that the plant responses to fungal VCs are light-dependent. Transcriptomic analyses of Arabidopsis leaves exposed to A. alternata VCs revealed changes in the expression of light- and CK-responsive genes involved in photosynthesis, growth and flowering. Notably, many genes differentially expressed in plants treated with fungal VCs were also differentially expressed in plants exposed to VCs emitted by the plant growth promoting rhizobacterium Bacillus subtilis GB03, suggesting that plants react to microbial VCs through highly conserved regulatory mechanisms.

Pinus pinaster Ait. is a conifer widely distributed through Spain. It is considered one of the major forest species because of wood and resin production. This fact explains the efforts of companies for capturing and enhancing the genetic gain in breeding programs. Micropropagation is a useful strategy to obtain identical plants from selected material, but nowadays, organogenesis protocols are the unique techniques to obtain clonal plants from a high number of elite genotypes. In this work, an improved micropropagation protocol for maritime pine using zygotic embryos as initial explant was developed. To optimize the protocol, the study of culture medium and exogenously applied cytokinins was needed. Embryos cultured on 1/2 DCR as basal medium exhibited high organogenic responses during induction, elongation, and rooting periods. Explants cultured on medium with 2.2 µM meta-topolin showed the highest values in all parameters analyzed along the different micropropagation phases, from induction to ac...

This paper describes a micropropagation protocol for in vitro propagation of mature Stone Pine trees. Axillary bud development was achieved by culturing bud explants in media containing various cytokinins. Experiments were conducted to test the effect of asepsis conditions, type and concentration of cytokinin and rooting protocol. Four cytokinins were tested, namely, benzyladenine, meta-topolin, N-benzyl-9-(2-tetrahydropyranyl)-adenine and thidiazuron (TDZ) of which TDZ gave the best results, as 59% shoot development was obtained following the application of 1 μM TDZ to the culture medium. The shoot development was significantly influenced by the genotype of the tree, but was effective in explants from all 20 genotypes used in the trial. In vitro rooting was, however, difficult to achieve and could only be induced at low rates. This protocol represents the first successful biotechnological approach to the micropropagation of adult Pinus pinea trees.

Plant hormones cytokinins regulate various aspects of plant growth and development. For their positive effects on branching, delaying of senescence, nutrient remobilisation, flower and seed set control they became interesting substances in search for potential agrochemicals. From the 1970' of the last century exogenous application of cytokinins have been tested in field conditions to improve yield traits of world-wide important crops such as wheat, rice, maize, barley, and soybean. Despite the extensive testing summarized in this work, so far cytokinins haven't found their stable place among commercialized plant growth regulators, mainly due to the complexity of their effects. Here we bring an overview of the outcomes obtained in pot and field experiments using cytokinin exogenous treatments, summarize the ways of application and point to the affected traits in various field crops, vegetables, cotton and fruit trees. Further, we present here outcomes of field trials performed with a derivative of N(6)-benzyladenine, 2-chloro-6-(3-methoxybenzyl)aminopurine, in spring barley and winter wheat. The effect on yield forming traits such as number of tillers, grains per ear, number of ears and the final yield was evaluated and compared after spraying of the both crops in different phenological stages.

Bahaji, Abdellatif et al., Phosphoglucose isomerase (PGI) catalyzes the reversible isomerization of glucose-6-phosphate and fructose-6-phosphate. It is involved in glycolysis and in the regeneration of glucose-6-P molecules in the oxidative pentose phosphate pathway (OPPP). In chloroplasts of illuminated mesophyll cells PGI also connects the Calvin-Benson cycle with the starch biosynthetic pathway. In this work we isolated pgi1-3, a mutant totally lacking pPGI activity as a consequence of aberrant intron splicing of the pPGI encoding gene, PGI1. Starch content in pgi1-3 source leaves was ca. 10-15% of that of wild type (WT) leaves, which was similar to that of leaves of pgi1-2, a T-DNA insertion pPGI null mutant. Starch deficiency of pgi1 leaves could be reverted by the introduction of a sex1 null mutation impeding β-amylolytic starch breakdown. Although previous studies showed that starch granules of pgi1-2 leaves are restricted to both bundle sheath cells adjacent to the mesophyll and stomata guard cells, microscopy analyses carried out in this work revealed the presence of starch granules in the chloroplasts of pgi1-2 and pgi1-3 mesophyll cells. RT-PCR analyses showed high expression levels of plastidic and extra-plastidic β-amylase encoding genes in pgi1 leaves, which was accompanied by increased β-amylase activity. Both pgi1-2 and pgi1-3 mutants displayed slow growth and reduced photosynthetic capacity phenotypes even under continuous light conditions. Metabolic analyses revealed that the adenylate energy charge and the NAD(P)H/NAD(P) ratios in pgi1 leaves were lower than those of WT leaves. These analyses also revealed that the content of plastidic 2-C-methyl-D-erythritol 4-phosphate (MEP)-pathway derived cytokinins (CKs) in pgi1 leaves were exceedingly lower than in WT leaves. Noteworthy, exogenous application of CKs largely reverted the low starch content phenotype of pgi1 leaves. The overall data show that pPGI is an important determinant of photosynthesis, energy status, growth and starch accumulation in mesophyll cells likely as a consequence of its involvement in the production of OPPP/glycolysis intermediates necessary for the synthesis of plastidic MEP-pathway derived hormones such as CKs., This work was partially supported by the Comisión Interministerial de Ciencia y Tecnología and Fondo Europeo de Desarrollo Regional (Spain) [grant numbers BIO2010-18239 and BIO2013-C2-1-P] and by the Government of Navarra [grant number IIM010491.RI1], the Ministry of Education, Youth and Sports of the Czech Republich [Grant L01204 from the National Program of Sustainability] and the European Social Fund and the state budget of the Czech Republic [project POST-UP, reg. No. CZ.1.07/2.3.00/30.0004]. AMS-L acknowledges a predoctoral fellowship from the Spanish Ministry of Science and Innovation. MB acknowledges a post-doctoral fellowship from the Public University of Navarra.