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The Peppermint Initiative seeks to inform the standardisation of breath analysis methods. Five Peppermint Experiments with gas chromatography-ion mobility spectrometry (GC-IMS), operating in the positive mode with a tritium 3H 5.68 keV, 370 MBq ionisation source, were undertaken to provide benchmark Peppermint Washout data for this technique, to support its use in breath-testing, analysis, and research. Headspace analysis of a peppermint-oil capsule by GC-IMS with on-column injection (0.5 cm3) identified 12 IMS responsive compounds, of which the four most abundant were: eucalyptol; β-pinene; α-pinene; and limonene. Elevated concentrations of these four compounds were identified in exhaled-breath following ingestion of a peppermint-oil capsule. An unidentified compound attributed as a volatile catabolite of peppermint-oil was also observed. The most intense exhaled peppermint-oil component was eucalyptol, which was selected as a peppermint marker for benchmarking GC-IMS. Twenty-five washout experiments monitored levels of exhaled eucalyptol, by GC-IMS with on-column injection (0.5 cm3), at t = 0 min, and then at t + 60, t + 90, t + 165, t + 285 and t + 360 min from ingestion of a peppermint capsule resulting in 148 peppermint breath analyses. Additionally, the Peppermint Washout data was used to evaluate clinical deployments with a further five washout tests run in clinical settings generating an additional 35 breath samples. Regression analysis yielded an average extrapolated time taken for exhaled eucalyptol levels to return to baseline values to be 429 ± 62 min (±95% confidence-interval). The benchmark value was assigned to the lower 95% confidence-interval, 367 min. Further evaluation of the data indicated that the maximum number of volatile organic compounds discernible from a 0.5 cm3 breath sample was 69, while the use of an in-line biofilter appeared to reduce this to 34.

Black fungi of the order Chaetothyriales are grown by many tropical plant-mutualistic ants as small so-called “patches” in their nests, which are located inside hollow structures provided by the host plant (“domatia”). These fungi are introduced and fostered by the ants, indicating that they are important for the colony. As several species of Chaetothyriales tolerate, adsorb, and metabolize toxic volatiles, we investigated the composition of volatile organic compounds (VOCs) of selected domatia in the Azteca/Cecropia ant-plant mutualism. Concentrations of VOCs in ant-inhabited domatia, empty domatia, and background air were compared. In total, 211 compounds belonging to 19 chemical families were identified. Ant-inhabited domatia were dominated by ketones with 2-heptanone, a well-known ant alarm semiochemical, as the most abundant volatile. Empty domatia were characterized by relatively high concentrations of the monoterpenes d-limonene, p-cymene and β-phellandrene, as well as the heterocyclic sulphur-containing compound, benzothiazole. These compounds have biocidal properties and are primarily biosynthesized by plants as a defense mechanism. Interestingly, most of the latter compounds were present at lower concentrations in ant inhabited domatia than in non-colonized ones. We suggest that Chaetothyriales may play a role in reducing the VOCs, underlining that the mutualistic nature of these fungi as VOCs accumulation might be detrimental for the ants, especially the larvae.

Dual-comb spectroscopy can provide broad spectral bandwidth and high spectral resolution in a short acquisition time, enabling time-resolved measurements. Specifically, spectroscopy in the mid-infrared wavelength range is of particular interest, since most of the molecules have their strongest rotational-vibrational transitions in this "fingerprint" region. Here we report time-resolved mid-infrared dual-comb spectroscopy for the first time, covering ~300 nm bandwidth around 3.3 {\mu}m with 6 GHz spectral resolution and 20 {\mu}s temporal resolution. As a demonstration, we study a CH4/He gas mixture in an electric discharge, while the discharge is modulated between dark and glow regimes. We simultaneously monitor the production of C2H6 and the vibrational excitation of CH4 molecules, observing the dynamics of both processes. This approach to broadband, high-resolution, and time-resolved mid-infrared spectroscopy provides a new tool for monitoring the kinetics of fast chemical reactions, with potential applications in various fields such as physical chemistry and plasma/combustion analysis., Comment: 21 page, 6 figures

A wide range of volatile organic compounds (VOCs) is released during plant-pathogen interactions both by the pathogens and the hosts. Some of these VOCs may have an effect on plant signal transduction pathways, thus contributing to pathogenicity or plant defence responses. Strains of the virulent biovar 3 of Pseudomonas syringae pv. actinidiae emit ethylene, both from axenic cultures in a growth medium obtained from Actinidia deliciosa leaf extract, and from infected plants. Accordingly, the infection stimulates the ethylene signal cascade in the host. Other compounds, either emitted by the bacterium (such as 1-undecene, linear ketones and benzyl alcohol), or strongly induced in plants after infection, such as decane, may present a biological activity. Healthy plants react to VOCs from infected neighbours, by activating the phenylpropanoid pathway and several PR genes. However, after the infection, P. syringae pv. actinidiae is able to selectively depress the induced plant defences, possibly by stimulating the oxylipin pathway. Ethylene and oxylipin derivatives, such as jasmonates, may counteract salicylic acid-mediated defences, thus allowing bacterial growth in the host plant. Our research demonstrates that VOCs from infected plants may promote both plant defences and disease development. The further characterization of the signalling pathways involved in such mechanisms may result useful to design plant protection strategies.

Nitrate and ammonia deferentially modulate primary metabolism during the hypersensitive response in tobacco. In this study, tobacco RNAi lines with low nitrite reductase (NiRr) levels were used to investigate the roles of nitrite and nitric oxide (NO) in this process. The lines accumulate NO2–, with increased NO generation, but allow sufficient reduction to NH4+ to maintain plant viability. For wild-type (WT) and NiRr plants grown with NO3–, inoculation with the non-host biotrophic pathogen Pseudomonas syringae pv. phaseolicola induced an accumulation of nitrite and NO, together with a hypersensitive response (HR) that resulted in decreased bacterial growth, increased electrolyte leakage, and enhanced pathogen resistance gene expression. These responses were greater with increases in NO or NO2– levels in NiRr plants than in the WT under NO3– nutrition. In contrast, WT and NiRr plants grown with NH4+ exhibited compromised resistance. A metabolomic analysis detected 141 metabolites whose abundance was differentially changed as a result of exposure to the pathogen and in response to accumulation of NO or NO2–. Of these, 13 were involved in primary metabolism and most were linked to amino acid and energy metabolism. HR-associated changes in metabolism that are often linked with primary nitrate assimilation may therefore be influenced by nitrite and NO production.

A mid-infrared trace gas detection system based on off-axis integrated cavity output spectroscopy (OA-ICOS) is demonstrated for accurate and sensitive detection of N2O in combination with a continuous wave external-cavity quantum cascade laser (EC-QCL) working around 7.7 µm. A 13-times improvement in signal-to-noise ratio is achieved using a re-injection mirror and a minimum detection limit of 70 ppbv in less than 10 s averaging time is achieved, which yields a noise-equivalent absorption sensitivity (NEAS) of 6×10−9 cm−1 Hz−1/2. For comparison, a compact multipass cell is deployed to measure the same absorption line of N2O using wavelength modulation spectroscopy with second harmonic detection (WMS-2f). An enhancement factor of 20 in comparison to direct absorption spectroscopy (DAS) is achieved, yielding a minimum detection limit of 15 ppbv in less than 10 s averaging and a NEAS of 1×10−9 cm−1 Hz−1/2. A comprehensive comparison between the two systems is carried out in terms of residual amplitude noise (RAM), linearity, long-term stability, detection limit, spectral fitting, reproducibility, and background variations. The proposed sensor based on OA-ICOS is potentially advantageous for trace gas sensing in outdoor applications and harsh environments due to its robustness and flexibility of alignment.

Ethylene interacts with other plant hormones to modulate many aspects of plant metabolism, including defence and stomata regulation. Therefore, its manipulation may allow plant pathogens to overcome the host’s immune responses. This work investigates the role of ethylene as a virulence factor for Pseudomonas syringae pv. actinidiae (Psa), the aetiological agent of the bacterial canker of kiwifruit. The pandemic, highly virulent biovar of this pathogen produces ethylene, whereas the biovars isolated in Japan and Korea do not. Ethylene production is modulated in planta by light/dark cycle. Exogenous ethylene application stimulates bacterial virulence, and restricts or increases host colonisation if performed before or after inoculation, respectively. The deletion of a gene, unrelated to known bacterial biosynthetic pathways and putatively encoding for an oxidoreductase, abolishes ethylene production and reduces the pathogen growth rate in planta. Ethylene production by Psa may be a recently and independently evolved virulence trait in the arms race against the host. Plant- and pathogen-derived ethylene may concur in the activation/suppression of immune responses, in the chemotaxis toward a suitable entry point, or in the endophytic colonisation.

A major challenge for breath research is the lack of standardization in sampling and analysis. To address this, a test that utilizes a standardized intervention and a defined study protocol has been proposed to explore disparities in breath research across different analytical platforms and to provide benchmark values for comparison. Specifically, the Peppermint Experiment involves the targeted analysis in exhaled breath of volatile constituents of peppermint oil after ingestion of the encapsulated oil. Data from the Peppermint Experiment performed by proton transfer reaction mass spectrometry (PTR-MS) and selected ion flow tube mass spectrometry (SIFT-MS) are presented and discussed herein, including the product ions associated with the key peppermint volatiles, namely limonene, alpha- and beta-pinene, 1,8-cineole, menthol, menthone and menthofuran. The breath washout profiles of these compounds from 65 individuals were collected, comprising datasets from five PTR-MS and two SIFT-MS instruments. The washout profiles of these volatiles were evaluated by comparing the log-fold change over time of the product ion intensities associated with each volatile. Benchmark values were calculated from the lower 95% confidence interval of the linear time-to-washout regression analysis for all datasets combined. Benchmark washout values from PTR-MS analysis were 353 min for the sum of monoterpenes and 1,8-cineole (identical product ions), 173 min for menthol, 330 min for menthofuran, and 218 min for menthone; from SIFT-MS analysis values were 228 min for the sum of monoterpenes, 281 min for the sum of monoterpenes and 1,8-cineole, and 370 min for menthone plus 1,8-cineole. Large inter- and intra-dataset variations were observed, whereby the latter suggests that biological variability plays a key role in how the compounds are absorbed, metabolized and excreted from the body via breath. This variability seems large compared to the influence of sampling and analytical procedures, but further investigations are recommended to clarify the effects of these factors. Marie Skodowska-Curie Actions Innovative Training Network: Ion-Molecule Processes for Analytical Chemistry Technologies (IMPACT) through the European Commission's HORIZON 2020 Programme [674911]; UK's National Physical Laboratory

Recent research discovered that airborne microplastics pollution can reach very remote areas. However, analysis of nanoplastics in environmental samples remains challenging, mostly due to technical and methodological issues. A new method for chemical characterisation of nanoplastics based on TD-PTR-MS has been recently introduced. The detection limit of

The production of volatile organic compounds (VOCs) during programmed cell death (PCD) is still insufficiently studied and their implication in the process is not well understood. The present study demonstrates that the release of VOSCs with presumed antioxidant capacity (methanethiol, dimethylsulfide and dimethyldisulfide) accompanies the cell death in chemical-stressed tobacco BY-2 suspension cultured cells. The cells were exposed to cell death inducers of biotic nature mastoparan (MP, wasp venom) and camptothecin (CPT, alkaloid), and to the abiotic stress agent CdSO4. The VOCs emission was monitored by proton-transfer reaction mass spectrometry (PTR-MS). The three chemicals induced PCD expressing apoptotic-like phenotype. The identified VOSCs were emitted in response to MP and CPT but not in presence of Cd. The VOSCs production occurred within few hours after the administration of the elicitors, peaked up when 20-50 % of the cells were dead and further levelled off with cell death advancement. This suggests that VOSCs with antioxidant activity may contribute to alleviation of cell death-associated oxidative stress at medium severity of cell death in response to the stress factors of biotic origin. The findings provide novel information about cell death defence mechanisms in chemical-challenged BY-2 cells and show that PCD related VOSCs synthesis depends on the type of inducer.

We present a multi-species trace gas sensor based on a high-repetition-rate mid-infrared supercontinuum source, in combination with a 30 m multipass absorption cell, and a scanning grating spectrometer. The output of the spectrometer is demodulated by a digital lock-in amplifier, referenced to the repetition rate of the supercontinuum source. This improved the detection sensitivity of the system by a factor 5, as compared to direct baseband operation. The spectrometer provides a spectral coverage of 950 cm−1 (between 2.85-3.90 µm) with a resolution of 2.5 cm−1 in 100 ms. It can achieve noise equivalent detection limits in the order of 100 ppbv Hz−1/2 for various hydrocarbons, alcohols, and aldehydes.

Sampling of volatile organic compounds (VOCs) has shown promise for detection of a range of diseases but results have proved hard to replicate due to a lack of standardization. In this work we introduce the 'Peppermint Initiative'. The initiative seeks to disseminate a standardized experiment that allows comparison of breath sampling and data analysis methods. Further, it seeks to share a set of benchmark values for the measurement of VOCs in breath. Pilot data are presented to illustrate the standardized approach to the interpretation of results obtained from the Peppermint experiment. This pilot study was conducted to determine the washout profile of peppermint compounds in breath, identify appropriate sampling time points, and formalise the data analysis. Five and ten participants were recruited to undertake a standardized intervention by ingesting a peppermint oil capsule that engenders a predictable and controlled change in the VOC profile in exhaled breath. After collecting a pre-ingestion breath sample, five further samples are taken at 2, 4, 6, 8, and 10 h after ingestion. Samples were analysed using ion mobility spectrometry coupled to multi-capillary column and thermal desorption gas chromatography mass spectrometry. A regression analysis of the washout data was used to determine sampling times for the final peppermint protocol, and the time for the compound measurement to return to baseline levels was selected as a benchmark value. A measure of the quality of the data generated from a given technique is proposed by comparing data fidelity. This study protocol has been used for all subsequent measurements by the Peppermint Consortium (16 partners from seven countries). So far 1200 breath samples from 200 participants using a range of sampling and analytical techniques have been collected. The data from the consortium will be disseminated in subsequent technical notes focussing on results from individual platforms. The Marie Sklodowska-Curie Actions Innovative Training Network: Ion-Molecule Processes for Analytical Chemistry Technologies (IMPACT) (www.impact-h2020itn.com), which has supported BH, MM and GP as Early Stage Researchers through the European Commission's HORIZON 2020 Programme under Grant Agreement Number 674911. The EMBER consortium for support in the dissemination of the Peppermint Initiative; researchers involved in the application of the protocol to specific techniques (to be published in due course) Abigail Joyce, Amisha Manek, Bellagambi Francesca Giuseppa, Bonsignori Alessio, Ghimenti Silvia and Biagini Denise, Liesl Carr, Luke Bryant and Rebecca Cordell. MW and SJF are supported by the NIHR Manchester Biomedical Research Centre. The group from Li`ege University is supported by the FNRS through the EOS and the FRIA initiatives. Ruszkiewicz, DM (, orresponding author), Loughborough Univ, Ctr Analyt Sci, Dept Chem, Loughborough, Leics, England ; Univ Birmingham, Sch Phys & Astron, Mol Phys Grp, Birmingham, W Midlands, England. D.M.Ruszkiewicz2@lboro.ac.uk

International audience; Agricultural activities highly contribute to atmospheric pollution, but the diversity and the magnitude of their emissions are still subject to large uncertainties. A field measurement campaign was conducted to characterize gaseous and particulate emissions from an experimental farm in France containing a sheep pen and a dairy stable. During the campaign, more than four hundred volatile organic compounds (VOCs) were characterized using an original combination of online and off-line measurements. Carbon dioxide (CO2) and ammonia (NH3) were the most concentrated compounds inside the buildings, followed by methanol, acetic acid and acetaldehyde. A CO2 mass balance model was used to estimate NH3 and VOC emission rates. To our knowledge, this study constitutes the first evaluation of emission rates for most of the identified VOCs. The measurements show that the dairy stable emitted more VOCs than the sheep pen. Despite strong VOC and NH3 emissions, the chemical composition of particles indicates that gaseous farm emissions do not affect the loading of fine particles inside the farm and is mainly explained by the low residence time inside the buildings. The experimental dataset obtained in this work will help to improve emissions inventories for agricultural activities.

Contains fulltext : 181450.pdf (Publisher’s version ) (Closed access) Herbivore-induced plant volatiles (HIPVs) serve as specific cues to higher trophic levels. Novel, exotic herbivores entering native foodwebs may disrupt the infochemical network as a result of changes in HIPV profiles. Here, we analysed HIPV blends of native Brassica rapa plants infested with one of 10 herbivore species with different coexistence histories, diet breadths and feeding modes. Partial least squares (PLS) models were fitted to assess whether HIPV blends emitted by Dutch B. rapa differ between native and exotic herbivores, between specialists and generalists, and between piercing-sucking and chewing herbivores. These models were used to predict the status of two additional herbivores. We found that HIPV blends predicted the evolutionary history, diet breadth and feeding mode of the herbivore with an accuracy of 80% or higher. Based on the HIPVs, the PLS models reliably predicted that Trichoplusia ni and Spodoptera exigua are perceived as exotic, leaf-chewing generalists by Dutch B. rapa plants. These results indicate that there are consistent and predictable differences in HIPV blends depending on global herbivore characteristics, including coexistence history. Consequently, native organisms may be able to rapidly adapt to potentially disruptive effects of exotic herbivores on the infochemical network. 13 p.

In nature, plants are frequently subjected to multiple biotic and abiotic stresses, resulting in a convergence of adaptive responses. We hypothesised that hormonal signalling regulating defences to different herbivores may interact with drought responses, causing distinct resistance phenotypes. To test this, we studied the hormonal and transcriptomic responses of Solanum dulcamara subjected to drought and herbivory by the generalist Spodoptera exigua (beet armyworm, BAW) or the specialist Leptinotarsa decemlineata (Colorado potato beetle, CPB). Bioassays showed that the performance of BAW, but not CPB, decreased on plants under drought compared to controls. While drought did not alter BAW-induced hormonal responses, it enhanced the CPB-induced accumulation of jasmonic acid and salicylic acid (SA), and suppressed ethylene (ET) emission. Microarray analyses showed that under drought, BAW herbivory enhanced several herbivore-induced responses, including cell-wall remodelling and the metabolism of carbohydrates, lipids, and secondary metabolites. In contrast, CPB herbivory enhanced several photosynthesis-related and pathogen responses in drought-stressed plants. This may divert resources away from defence production and increase leaf nutritive value. In conclusion, while BAW suffers from the drought-enhanced defences, CPB may benefit from the effects of enhanced SA and reduced ET signalling. This suggests that the fine-tuned interaction between the plant and its specialist herbivore is sustained under drought.

In the field of laser-based absorption spectroscopy, off-axis integrated cavity output spectroscopy is considered to be a sensitive and robust method, employing a simple optical design. However, one of the major drawbacks of non-mode-matched cavities combined with highly reflective mirrors (>99.98%) is its low output intensity. Here, we systematically investigate the increase in cavity output intensity, using a third re-injection mirror before the absorption cavity. The presented design not only enables high transmission power but also retains a long effective path length. To investigate the intensity enhancement, we used a CO2 absorption line in the near-IR wavelength region at 6240.10 cm−1. In agreement with our simulation model, we achieved an intensity enhancement factor of 38. We achieved a noise equivalent absorption sensitivity to 1.6×10−8 cm−1 Hz−1/2, which is no longer limited by the detectivity of the detector.

Volatile organic compounds emitted during the infection of apple (Malus pumila var. domestica) plants by Erwinia amylovora or Pseudomonas syringae pv. syringae were studied by gas chromatography-mass spectrometry and proton transfer reaction-mass spectrometry, and used to treat uninfected plants. Infected plants showed a disease-specific emission of volatile organic compounds, including several bio-active compounds, such as hexenal isomers and 2,3-butanediol. Leaf growth promotion and a higher resistance to the pathogen, expressed as a lower bacterial growth and migration in plant tissues, were detected in plants exposed to volatile compounds from E. amylovora-infected plants. Transcriptional analysis revealed the activation of salicylic acid synthesis and signal transduction in healthy plants exposed to volatiles produced by E. amylovora-infected neighbour plants. In contrast, in the same plants, salicylic acid-dependent responses were repressed after infection, whereas oxylipin metabolism was activated. These results clarify some metabolic and ecological aspects of the pathogenic adaptation of E. amylovora to its host.

We present a fully integrated and transportable multi-species trace gas sensor based on a mid-infrared (MIR) supercontinuum light source. The high brightness (surpassing synchrotron) and ultra-broad spectral bandwidth (2&ndash, 4 &mu, m) of this light source allows simultaneous detection of multiple broadband absorbing gas species. High sensitivity in the sub-ppmv level has been achieved by utilizing an astigmatic multipass cell. A grating-based spectrometer at a scanning rate of 20 Hz is developed employing a balanced detection scheme. A multi-component global fitting algorithm is implemented into a central LabVIEW program to perform real-time data analysis. The obtained concentration values are validated by the standard gas chromatography mass spectrometry (GC-MS) method. Field application of the sensor for quality control of stored fruits at a small scale is demonstrated, involving the detection of ethylene, ethanol, ethyl acetate, acetaldehyde, methanol, acetone, and water simultaneously. The sensor also shows promising potentials for other applications, such as environmental monitoring and biomedical research.

This study addressed the role of lipid-derived factors together with ethylene and ROS in programmed cell death (PCD) signaling in tobacco BY-2 suspension cultured cells. The cells were exposed to the chemical stress agents mastoparan (MP) and ethanol (EtOH). MP is an activator of membrane-associated heterotrimeric G-proteins and downstream phospholipids-dependent processes; EtOH is suggested to affect lipid-associated pathways. The effects of MP and EtOH were compared to cell death in response to CdSO4. All applied chemicals appeared potent cell death inducers. Ethylene and lipid signaling were found instrumental in chemical-induced cell death, presumably in conjunction with ROS. Cadmium and MP induced cell death of apoptotic-like phenotype. Lower EtOH concentrations (1–2%) induced vacuolar cell death associated with autophagy-associated formation of lysosome-like acidic organelles in part of the cells; in other cells in the same suspension apoptotic-like features were observed. At higher EtOH concentration (3%) the dead cells expressed exclusively apoptotic-like morphology. The results suggest that phospholipase D- and phospholipase C-derived phosphatidic acid triggers ROS generation that is responsible for the observed apoptotic-like PCD. Vacuolar cell death in EtOH-treated cells appeared dependent on phospholipase C - phosphatidylinositol-3-kinase-related pathway. The obtained results indicate that depending on the inducer and stress severity, similar or distinct regulatory pathways can be activated, and the signals may interact in the transmission of the cell death message.

Ethylene interacts with other plant hormones to modulate many aspects of plant metabolism, including defence and stomata regulation. Therefore, its manipulation may allow plant pathogens to overcome the host’s immune responses. This work investigates the role of ethylene as a virulence factor for Pseudomonas syringae pv. actinidiae (Psa), the aetiological agent of the bacterial canker of kiwifruit. The pandemic, highly virulent biovar of this pathogen produces ethylene, whereas the biovars isolated in Japan and Korea do not. Ethylene production is modulated in planta by light/dark cycle. Exogenous ethylene application stimulates bacterial virulence, and restricts or increases host colonisation if performed before or after inoculation, respectively. The deletion of a gene, unrelated to known bacterial biosynthetic pathways and putatively encoding for an oxidoreductase, abolishes ethylene production and reduces the pathogen growth rate in planta. Ethylene production by Psa may be a recently and independently evolved virulence trait in the arms race against the host. Plant- and pathogen-derived ethylene may concur in the activation/suppression of immune responses, in the chemotaxis toward a suitable entry point, or in the endophytic colonisation.

Ralstonia solanacearum (Rs) and Clavibacter michiganensis subsp. sepedonicus (Cms) are the bacterial causal agents of potato brown and ring rot, respectively, and are included in the A2 list of quarantine pathogens in Europe. Identification by GC-MS analysis of volatile organic compounds from Rs or Cms cultured on different nutrient media was performed. GC-MS and PTR-MS analysis were carried out also on unwounded potato tubers infected with the same pathogens. Infected tubers were produced by experimental inoculations of the plants. In in vitro experiments, Rs or Cms emitted volatile compounds, part of which were specific disease markers of potato (2-propanol and 3-methylbutanoic acid), mainly originating from bacterial metabolism (i.e., amino acid degradation, carbohydrate and fatty acid oxidation). In potato tubers, pathogen metabolism modified the volatile compound pattern emitted from healthy samples. Both bacteria seem to accelerate metabolic processes ongoing in potatoes and, in the case of Rs, disease markers (1-hepten-3-ol, 3,6-dimethyl-3-octanone, 3-ethyl-3-methylpentane, 1-chloroctane, and benzothiazole) were identified.

An original approach to develop sustainable agriculture with less nitrogen fertilizer inputs is to tackle the cross-talk between nitrogen nutrition and plant growth regulators. In particular the gaseous hormone, ethylene, is a prime target for that purpose. The variation of ethylene production in natural accessions of the model species Arabidopsis thaliana was explored in response to the nitrate supply. Ethylene was measured with a laser-based photoacoustic detector. First, experimental conditions were established with Columbia-0 (Col-0) accession, which was grown in vitro on horizontal plates across a range of five nitrate concentrations (0.5, 1, 2.5, 5 and 10 mM). The concentrations of 1 and 10 mM nitrate were retained for further characterization. Along with a decrease of total dry biomass and higher biomass allocation to the roots, the ethylene production was 50% more important at 1 mM than at 10 mM nitrate. The total transcript levels of 1-AMINOCYCLOPROPANE-1-CARBOXYLIC ACID SYNTHASES (ACS) in roots and those of ACC OXIDASES (ACO) in shoots increased by 100% between the same treatments. This was mainly due to higher transcript levels of ACS6 and of ACO2 and ACO4. The assumption was that during nitrogen deficiency, the greater biomass allocation in favor of the roots was controlled by ethylene being released in the shoots after conversion of ACC originating from the roots. Second, biomass and ethylene productions were measured in twenty accessions. Across all accessions, the total dry biomass and ethylene production were correlated negatively at 1 mM but positively at 10 mM nitrate. Furthermore, polymorphism was surveyed in ACC and ethylene biosynthesis genes and gene products among accessions. Very few substitutions modifying the amino acids properties in conserved motifs of the enzymes were found in the accessions. Natural variation of ethylene production could be further explored to improve Nitrogen Use Efficiency, in particular by manipulating features like the biomass production and the timing of senescence upon nitrogen limitation., SCOPUS: ar.j, info:eu-repo/semantics/published

In nature, plants are frequently subjected to multiple biotic and abiotic stresses, resulting in a convergence of adaptive responses. We hypothesised that hormonal signalling regulating defences to different herbivores may interact with drought responses, causing distinct resistance phenotypes. To test this, we studied the hormonal and transcriptomic responses of Solanum dulcamara subjected to drought and herbivory by the generalist Spodoptera exigua (beet armyworm; BAW) or the specialist Leptinotarsa decemlineata (Colorado potato beetle; CPB). Bioassays showed that the performance of BAW, but not CPB, decreased on plants under drought compared to controls. While drought did not alter BAW-induced hormonal responses, it enhanced the CPB-induced accumulation of jasmonic acid and salicylic acid (SA), and suppressed ethylene (ET) emission. Microarray analyses showed that under drought, BAW herbivory enhanced several herbivore-induced responses, including cell-wall remodelling and the metabolism of carbohydrates, lipids, and secondary metabolites. In contrast, CPB herbivory enhanced several photosynthesis-related and pathogen responses in drought-stressed plants. This may divert resources away from defence production and increase leaf nutritive value. In conclusion, while BAW suffers from the drought-enhanced defences, CPB may benefit from the effects of enhanced SA and reduced ET signalling. This suggests that the fine-tuned interaction between the plant and its specialist herbivore is sustained under drought.

We employed a single-mode, widely tunable (~300 cm&minus, 1) external-cavity quantum cascade laser operating around 8 &micro, m for broadband direct absorption spectroscopy and wavelength modulation spectroscopy where a modulation frequency of 50 kHz was employed with high modulation amplitudes of up to 10 GHz. Using a compact multipass cell, we measured the entire molecular absorption band of acetone at ~7.4 &micro, m with a spectral resolution of ~1 cm&minus, 1. In addition, to demonstrate the high modulation dynamic range of the laser, we performed direct absorption (DAS) and second harmonic wavelength modulation spectroscopy (WMS-2f) of the Q-branch peak of acetone molecular absorption band (HWHM ~10 GHz) near 1365 cm&minus, 1. With WMS-2f, a minimum detection limit of 15 ppbv in less than 10 s is achieved, which yields a noise equivalent absorption sensitivity of 1.9 &times, 10&minus, 8 cm&minus, 1 Hz&minus, 1/2.

Despite the established importance of nitric oxide (NO) in many physiological and molecular processes in plants, most methods for quantifying NO are open to criticism This reflects the differing methods either lacking specificity or sensitivity, or even from an undue dependence of results on experimental conditions (i.e., chemical concentrations, pH, etc.). In this chapter we describe a protocol to measure gaseous NO produced by a biological sample using quantum cascade laser (QCL)-based spectroscopy. This technique is based on absorption of the laser light by the NO molecules which have been passed from a biological sample into an optical s cell that is equipped with two mirrors placed at both ends. This design greatly increases the interaction path length with the NO molecules due to multiple reflections of the light coupled inside the cell. Thus, the method is able to provide online, in planta measurements of the dynamics of NO production, being highly selective and sensitive (down to ppbv levels;1 ppbv = part per billion by volume mixing ratio = 1:10

Detection and analysis of volatile compounds in exhaled breath represents an attractive tool for monitoring the metabolic status of a patient and disease diagnosis, since it is non-invasive and fast. Numerous studies have already demonstrated the benefit of breath analysis in clinical settings/applications and encouraged multidisciplinary research to reveal new insights regarding the origins, pathways, and pathophysiological roles of breath components. Many breath analysis methods are currently available to help explore these directions, ranging from mass spectrometry to laser-based spectroscopy and sensor arrays. This review presents an update of the current status of optical methods, using near and mid-infrared sources, for clinical breath gas analysis over the last decade and describes recent technological developments and their applications. The review includes: tunable diode laser absorption spectroscopy, cavity ring-down spectroscopy, integrated cavity output spectroscopy, cavity-enhanced absorption spectroscopy, photoacoustic spectroscopy, quartz-enhanced photoacoustic spectroscopy, and optical frequency comb spectroscopy. A SWOT analysis (strengths, weaknesses, opportunities, and threats) is presented that describes the laser-based techniques within the clinical framework of breath research and their appealing features for clinical use.

A novel approach to enhance the signal-to-noise ratio in off-axis integrated cavity output spectroscopy, centered around 8µm is demonstrated. This work exhibits cavity-enhanced spectroscopy using a compact size, with absorption sensitivity comparable to longer absorption cells.

A 3-D ray-tracing model is developed, to model reinjection in Off-Axis Cavity Enhanced Spectroscopy. Modelling and experiments demonstrate, at near-IR laser wavelengths, a 38-times enhancement, corresponding to a NEAS of 1.1 × 10−9 cm−1 Hz−1/2.

We present mid-infrared trace gas sensing based on a supercontinuum source combined with direct absorption spectroscopy and sum-frequency upconversion, showing broadband detection with a 10−4 Hz−1/2 noise equivalent absorbance sensitivity.

Despite the established importance of nitric oxide (NO) in many physiological and molecular processes in plants, most methods for quantifying NO are open to criticism This reflects the differing methods either lacking specificity or sensitivity, or even from an undue dependence of results on experimental conditions (i.e., chemical concentrations, pH, etc.). In this chapter we describe a protocol to measure gaseous NO produced by a biological sample using quantum cascade laser (QCL)-based spectroscopy. This technique is based on absorption of the laser light by the NO molecules which have been passed from a biological sample into an optical s cell that is equipped with two mirrors placed at both ends. This design greatly increases the interaction path length with the NO molecules due to multiple reflections of the light coupled inside the cell. Thus, the method is able to provide online, in planta measurements of the dynamics of NO production, being highly selective and sensitive (down to ppbv levels;1 ppbv = part per billion by volume mixing ratio = 1:10-9).

Proton transfer reaction mass spectrometry (PTR-MS) is employed as a highly sensitive detection method for trace gas analysis of sulfur compounds. The effects of drift tube humidity and the reduced electric field ( E / N value) were evaluated to measure traces of methanethiol, dimethyl sulfide (DMS) and dimethyl disulfide (DMDS) under optimal conditions. The three sulfur compounds showed a maximum intensity of the product ion signal at a low fraction of the protonated water clusters as compared to the total reagent ion signal. For methanethiol, DMS and DMDS the highest intensity was observed when the percentage branching ratio of H 3 O + ·H 2 O with total reagent ion signal was 3%, 5% and 3%, respectively. The methanethiol signal dropped sharply with an increase in H 3 O + ·H 2 O while DMS and DMDS showed a less strong dependence. This could be explained by their difference in proton affinity as compared to that of the water cluster (methanethiol 773 kJ mol −1 , DMS 830 kJ mol −1 , DMDS 815 kJ mol −1 , H 2 O·H 2 O >800 kJ mol −1 ). Taking into account the mean kinetic energy in the drift tube (19 kJ mol −1 ), the direct proton transfer reaction between DMS or DMDS and H 3 O + ·H 2 O is possible, in contrast to methanethiol. The PTR-MS was used to monitor trace gas emissions of methanethiol, DMS and DMDS emitted from Brassica rapa plants in real time. The concentrations of these sulfur compound emissions are in the part per billion range (ppbv) and optimization of the instrument for these compounds helps to measure the low concentrations emitted from control and infested plants. The plant roots were infested with Delia radicum larvae and all three sulfur compounds showed an immediate rise in the ion signal intensity after infestation, indicating that the plants were reacting after the roots were damaged by the insects. The sulfur compound emissions lasted for 30 h.

We demonstrate mid-infrared dual-comb spectroscopy with an optical parametric oscillator (OPO) toward real-time field measurement. A singly resonant OPO based on a MgO-doped periodically poled lithium niobate (PPLN) crystal is demonstrated. Chirped mirrors are used to compensate the dispersion caused by the optical cavity and the crystal. A low threshold of 17 mW has been achieved. The OPO source generates a tunable idler frequency comb between 2.7 and 4.7 μm. Dual-comb spectroscopy is achieved by coupling two identical Yb-fiber mode-locked lasers to this OPO with slightly different repetition frequencies. A measured absorption spectrum of methane is presented with a spectral bandwidth of $$300\,\hbox {cm}^{-1}$$ , giving an instrumental resolution of $$0.4\,\hbox {cm}^{-1}$$ . In addition, a second OPO containing two MgO-doped PPLN crystals in a singly resonant ring cavity is demonstrated. As such, this OPO generates two idler combs (average power up to 220 mW), covering a wavelength range between 2.7 and 4.2 μm, from which a mid-infrared dual-comb Fourier transform spectrometer is constructed. By detecting the heterodyned signal between the two idler combs, broadband spectra of molecular gases can be observed over a spectral bandwidth of more than $$350\,\hbox {cm}^{-1}$$ . This special cavity design allows the spectral resolution to be improved to $$0.2\,\hbox {cm}^{-1}$$ without locking the OPO cavity, indicating that this OPO represents an ideal high-power broadband mid-infrared source for real-time gas sensing.

Fruit and vegetables are regularly stored by consumers in the refrigerator at temperatures that may be well below the recommended storage temperatures. Apart from causing visible symptoms such as watery, sunken areas on the skin, chilling may also induce changes in fruit textural properties and flavour. The aim of this research was to investigate the effect of low temperature storage on tomato flavour and off-flavour production. To more closely mimic the real-consumer aroma perception while eating, in addition to the standard solid-phase microextraction gas chromatography-mass spectrometry (SPME/GC-MS) analysis, volatiles were also measured using a chewing device connected to a proton-transfer reaction-mass spectrometer (PTR-MS). Aroma volatiles were assessed in red ripe tomatoes of the cvs Cappricia RZ (round truss) and Amoroso RZ (cocktail truss) stored at refrigerator temperature (4 °C) and at higher temperatures (16 and 22 °C) for 20 days. The changes in aroma production were also monitored when the fruit was brought from room to refrigerator temperature and vice versa. After bringing the fruit from room to refrigerator temperature, the abundance of most volatiles was greatly reduced within 3 to 5 h, closely following the decrease in fruit temperature. When temperature was restored to room temperature following varying times of cold storage, the abundance of most volatiles increased again, but generally not to the original levels. Overall, the effects of low temperature storage on the decrease in volatile abundance were more pronounced in cv Cappricia RZ than in cv Amoroso RZ. On the contrary, the production of off flavours following prolonged cold storage was more pronounced in cv Amoroso RZ than in cv Cappricia RZ. Apart from changes in the overall abundance of the volatiles, marked changes in the volatile profile were observed in fruit stored for longer times in the cold and this may at least in part explain the negative effect of cold storage on overall tomato flavour.