Long-term measurements of chlorophyll a fluorescence using the JIP-test show that combined abiotic stresses influence the photosynthetic performance of the perennial ryegrass (Lolium perenne) in a managed temperate grassland.

Several experiments have highlighted the complexity of stress interactions involved in plant response. The impact in field conditions of combined environmental constraints on the mechanisms involved in plant photosynthetic response, however, remains understudied. In a long-term field study performed in a managed grassland, we investigated the photosynthetic apparatus response of the perennial ryegrass (Lolium perenne L.) to environmental constraints and its ability to recover and acclimatize. Frequent field measurements of chlorophyll a fluorescence (ChlF) were made in order to determine the photosynthetic performance response of a population of L. perenne. Strong midday declines in the maximum quantum yield of primary photochemistry (FV FM ) were observed in summer, when a combination of heat and high light intensity increased photosynthetic inhibition. During this period, increase in photosystem I (PSI) activity efficiency was also recorded, suggesting an increase in the photochemical pathway for de-excitation in summer. Strong climatic events (e.g. heat waves) were shown to reduce electron transport between photosystem II (PSII) and PSI. This reduction might have preserved the PSI from photo-oxidation. Periods of low soil moisture and high levels of sun irradiance increased PSII sensitivity to heat stress, suggesting increased susceptibility to combined environmental constraints. Despite the multiple inhibitions of photosynthetic functionality in summer, the L. perenne population showed increased PSII tolerance to environmental stresses in August. This might have been a response to earlier environmental constraints. It could also be linked to the selection and/or emergence of well-adapted individuals.

Influence of rhizobacterial volatiles on the root system architecture and the production and allocation of biomass in the model grass Brachypodium distachyon (L.) P. Beauv.

BACKGROUND: Plant growth-promoting rhizobacteria are increasingly being seen as a way of complementing conventional inputs in agricultural systems. The effects on their host plants are diverse and include volatile-mediated growth enhancement. This study sought to assess the effects of bacterial volatiles on the biomass production and root system architecture of the model grass Brachypodium distachyon (L.) Beauv. RESULTS: An in vitro experiment allowing plant-bacteria interaction throughout the gaseous phase without any physical contact was used to screen 19 bacterial strains for their growth-promotion ability over a 10-day co-cultivation period. Five groups of bacteria were defined and characterised based on their combined influence on biomass production and root system architecture. The observed effects ranged from unchanged to greatly increased biomass production coupled with increased root length and branching. Primary root length was increased only by the volatile compounds emitted by Enterobacter cloacae JM22 and Bacillus pumilus T4. Overall, the most significant results were obtained with Bacillus subtilis GB03, which induced an 81 % increase in total biomass, as well as enhancing total root length, total secondary root length and total adventitious root length by 88.5, 201.5 and 474.5 %, respectively. CONCLUSIONS: This study is the first report on bacterial volatile-mediated growth promotion of a grass plant. Contrasting modulations of biomass production coupled with changes in root system architecture were observed. Most of the strains that increased total plant biomass also modulated adventitious root growth. Under our screening conditions, total biomass production was strongly correlated with the length and branching of the root system components, except for primary root length. An analysis of the emission kinetics of the bacterial volatile compounds is being undertaken and should lead to the identification of the compounds responsible for the observed growth-promotion effects. Within the context of the inherent characteristics of our in vitro system, this paper identifies the next critical experimental steps and discusses them from both a fundamental and an applied perspective.

Reprogramming of fatty acid and oxylipin synthesis in rhizobacteria-induced systemic resistance in tomato.

The rhizobacterium Pseudomonas putida BTP1 stimulates induced systemic resistance (ISR) in tomato. A previous work showed that the resistance is associated in leaves with the induction of the first enzyme of the oxylipin pathway, the lipoxygenase (LOX), leading to a faster accumulation of its product, the free 13-hydroperoxy octadecatrienoic acid (13-HPOT), 2 days after Botrytis cinerea inoculation. In the present study, we further investigated the stimulation of the oxylipin pathway: metabolites and enzymes of the pathway were analyzed to understand the fate of the 13-HPOT in ISR. Actually the stimulation began upstream the LOX: free linolenic acid accumulated faster in P. putida BTP1-treated plants than in control. Downstream, the LOX products 13-fatty acid hydroperoxides esterified to galactolipids and phospholipids were more abundant in bacterized plants than in control before infection. These metabolites could constitute a pool that will be used after pathogen attack to produce free fungitoxic metabolites through the action of phospholipase A2, which is enhanced in bacterized plants upon infection. Enzymatic branches which can use as substrate the fatty acid hydroperoxides were differentially regulated in bacterized plants in comparison to control plants, so as to lead to the accumulation of the most fungitoxic compounds against B. cinerea. Our study, which is the first to demonstrate the accumulation of an esterified defense metabolite during rhizobacteria-mediated induced systemic resistance, showed that the oxylipin pathway is differentially regulated. It suggests that this allows the plant to prepare to a future infection, and to respond faster and in a more effective way to B. cinerea invasion.

EFSA's scientific activities and achievements on the risk assessment of genetically modified organisms (GMOs) during its first decade of existence: looking back and ahead.

Genetically modified organisms (GMOs) and derived food and feed products are subject to a risk analysis and regulatory approval before they can enter the market in the European Union (EU). In this risk analysis process, the role of the European Food Safety Authority (EFSA), which was created in 2002 in response to multiple food crises, is to independently assess and provide scientific advice to risk managers on any possible risks that the use of GMOs may pose to human and animal health and the environment. EFSA's scientific advice is elaborated by its GMO Panel with the scientific support of several working groups and EFSA's GMO Unit. This review presents EFSA's scientific activities and highlights its achievements on the risk assessment of GMOs for the first 10 years of its existence. Since 2002, EFSA has issued 69 scientific opinions on genetically modified (GM) plant market registration applications, of which 62 for import and processing for food and feed uses, six for cultivation and one for the use of pollen (as or in food), and 19 scientific opinions on applications for marketing products made with GM microorganisms. Several guidelines for the risk assessment of GM plants, GM microorganisms and GM animals, as well as on specific issues such as post-market environmental monitoring (PMEM) were elaborated. EFSA also provided scientific advice upon request of the European Commission on safeguard clause and emergency measures invoked by EU Member States, annual PMEM reports, the potential risks of new biotechnology-based plant breeding techniques, evaluations of previously assessed GMOs in the light of new scientific publications, and the use of antibiotic resistance marker genes in GM plants. Future challenges relevant to the risk assessment of GMOs are discussed. EFSA's risk assessments of GMO applications ensure that data are analysed and presented in a way that facilitates scientifically sound decisions that protect human and animal health and the environment.

Characterization of volatile organic compounds emitted by barley (Hordeum vulgare L.) roots and their attractiveness to wireworms.

Root volatile organic compounds (VOCs), their chemistry and ecological functions have garnered less attention than aboveground emitted plant VOCs. We report here on the identification of VOCs emitted by barley roots (Hordeum vulgare L.). Twenty nine VOCs were identified from isolated 21-d-old roots. The detection was dependent on the medium used for root cultivation. We identified 24 VOCs from 7-d-old roots when plants were cultivated on sterile Hoagland gelified medium, 33 when grown on sterile vermiculite, and 34 on non-sterile vermiculite. The major VOCs were fatty acid derived compounds, including hexanal, methyl hexanoate, (E)-hex-2-enal, 2-pentylfuran, pentan-1-ol, (Z)-2-(pentenyl)-furan, (Z)-pent-2-en-1-ol, hexan-1-ol, (Z)-hex-3-en-1-ol, (E)-hex-2-en-1-ol, oct-1-en-3-ol, 2-ethylhexan-1-ol (likely a contaminant), (E)-non-2-enal, octan-1-ol, (2E,6Z)-nona-2,6-dienal, methyl (E)-non-2-enoate, nonan-1-ol, (Z)-non-3-en-1-ol, (E)-non-2-en-1-ol, nona-3,6-dien-1-ol, and nona-2,6-dien-1-ol. In an olfactometer assay, wireworms (larvae of Agriotes sordidus Illiger, Coleoptera: Elateridae) were attracted to cues emanating from barley seedlings. We discuss the role of individual root volatiles or a blend of the root volatiles detected here and their interaction with CO2 for wireworm attraction.

Improvement of ylang-ylang essential oil characterization by GC×GC-TOFMS.

A single fraction of essential oil can often contain hundreds of compounds. Despite of the technical improvements and the enhanced selectivity currently offered by the state-of-the-art gas chromatography (GC) and mass spectrometry (MS) instruments, the complexity of essential oils is frequently underestimated. Comprehensive two-dimensional GC coupled to time-of-flight MS (GC×GC-TOFMS) was used to improve the chemical characterization of ylang-ylang essential oil fractions recently reported in a previous one-dimensional (1D) GC study. Based on both, the enhanced chromatographic separation and the mass spectral deconvolution, 161 individual compounds were identified and labeled as potentially characteristic analytes found in both low and high boiling fractions issued from distillation of mature ylang-ylang flowers. Compared to the most recent full GC-MS characterization, this represents 75 new compounds, essentially consisting of terpenes, terpenoid esters, and alcohols.

Comparative chemical and molecular variability of Cananga odorata (Lam.) Hook.f. & Thomson forma genuina (ylang-ylang) in the Western Indian Ocean Islands: implication for valorization.

Cananga odorata (Lam.) Hook.f. & Thomson forma genuina (Annonaceae) is a tropical tree, grown for the production of ylang-ylang essential oil, which is extracted from its fresh and mature flowers. Despite its economic and social importance, very little information is available on its variability and the possible factors causing it. Therefore, the relationship between the genetic structure, revealed by amplified fragment length polymorphism (AFLP), and the essential oil chemical composition, determined by GC/MS analysis, of ylang-ylang grown in semi-managed systems in three Indian Ocean islands (Grande Comore, Mayotte, and Madagascar) was investigated. Our results revealed a low genetic variation within plantations and contrasted situations between islands. Variations of the chemical composition could be observed within plantations and between islands. The genetic differentiation pattern did not match the observed pattern of chemical variability. Hence, the chemical variation could not be attributed to a genetic control. As Grande Comore, Madagascar, and Mayotte present different environmental and agronomic conditions, it can be concluded that the influence of these conditions on the ylang-ylang essential oil composition is consistent with the patterns observed. Finally, several strategies were proposed to valorize the chemical composition variations.

Developmental plasticity of Brachypodium distachyon in response to P deficiency: Modulation by inoculation with phosphate-solubilizing bacteria.

BACKGROUND: Mineral phosphorus (P) fertilizers must be used wisely in order to preserve rock phosphate, a limited and non-renewable resource. The use of bio-inoculants to improve soil nutrient availability and trigger an efficient plant response to nutrient deficiency is one potential strategy in the attempt to decrease P inputs in agriculture. METHOD: An in vitro co-cultivation system was used to study the response of Brachypodium distachyon to contrasted P supplies (soluble and poorly soluble forms of P) and inoculation with P solubilizing bacteria. Brachypodium's responses to P conditions and inoculation with bacteria were studied in terms of developmental plasticity and P use efficiency. RESULTS: Brachypodium showed plasticity in its biomass allocation pattern in response to variable P conditions, specifically by prioritizing root development over shoot productivity under poorly soluble P conditions. Despite the ability of the bacteria to solubilize P, shoot productivity was depressed in plants inoculated with bacteria, although the root system development was maintained. The negative impact of bacteria on biomass production in Brachypodium might be attributed to inadequate C supply to bacteria, an increased competition for P between both organisms under P-limiting conditions, or an accumulation of toxic bacterial metabolites in our cultivation system. Both P and inoculation treatments impacted root system morphology. The modulation of Brachypodium's developmental response to P supplies by P solubilizing bacteria did not lead to improved P use efficiency. CONCLUSION: Our results support the hypothesis that plastic responses of Brachypodium cultivated under P-limited conditions are modulated by P solubilizing bacteria. The considered experimental context impacts plant-bacteria interactions. Choosing experimental conditions as close as possible to real ones is important in the selection of P solubilizing bacteria. Both persistent homology and allometric analyses proved to be useful tools that should be considered when studying the impact of bio-inoculants on plant development in response to varying nutritional context.

Quantitative Structure-Activity Relationship of Humic-Like Biostimulants Derived From Agro-Industrial Byproducts and Energy Crops.

Humic-like substances (HLSs) isolated by alkaline oxidative hydrolysis from lignin-rich agro-industrial residues have been shown to exert biostimulant activity toward maize (Zea mays L.) germination and early growth. The definition of a quantitative structure-activity relationship (QSAR) between HLS and their bioactivity could be useful to predict their biological properties and tailor plant biostimulants for specific agronomic and industrial uses. Here, we created several projection on latent structure (PLS) regression by using published analytical data on the molecular composition of lignin-derived HLS obtained by both 13C-CPMAS-NMR spectra directly on samples and 31P-NMR spectra after derivatization of hydroxyl functions with a P-containing reagent (2-chloro-4,4,5,5-tetramethyl-1,3,2-dioxaphospholane). These spectral data were used to model the effect of HLS on the elongation of primary root, lateral seminal roots, total root apparatus, and coleoptile of maize. The 13C-CPMAS-NMR data suggested that methoxyl and aromatic moieties positively affected plant growth, while the carboxyl/esterified functions showed a negative impact on the overall seedling development. Alkyl C seems to promote Col elongation while concomitantly reducing that of the root system. Additionally, 31P-NMR-derived spectra revealed that the elongation of roots and Col were enhanced by the occurrence of aliphatic hydroxyl groups, and guaiacyl and p-Hydroxyphenyl lignin monomers. The PLS models based on raw dataset from 13C-CPMAS-NMR spectra explained more than 74% of the variance for the length of lateral seminal roots, total root system and coleoptile, while other parameters derived from 13C-CPMAS-NMR spectra, namely the Hydrophobicity and Hydrophilicity of materials were necessary to explain 83% of the variance of the primary root length. The results from 31P-NMR spectra explained the observed biological variance by 90, 96, 96, and 93% for the length of primary root, lateral seminal roots, total root system and coleoptile, respectively. This work shows that different NMR spectroscopy techniques can be used to build up PLS models which can predict the bioactivity of lignin-derived HLS toward early growth of maize plants. The established QSAR may also be exploited to enhance by chemical techniques the bioactive properties of HLS and enhance their plant stimulation capacity.

Attacks by a piercing-sucking insect (Myzus persicae Sultzer) or a chewing insect (Leptinotarsa decemlineata Say) on potato plants (Solanum tuberosum L.) induce differential changes in volatile compound release and oxylipin synthesis.

Plant defensive strategies bring into play blends of compounds dependent on the type of attacker and coming from different synthesis pathways. Interest in the field is mainly focused on volatile organic compounds (VOCs) and jasmonic acid (JA). By contrast, little is known about the oxidized polyunsaturated fatty acids (PUFAs), such as PUFA-hydroperoxides, PUFA-hydroxides, or PUFA-ketones. PUFA-hydroperoxides and their derivatives might be involved in stress response and show antimicrobial activities. Hydroperoxides are also precursors of JA and some volatile compounds. In this paper, the differential biochemical response of a plant against insects with distinct feeding behaviours is characterized not only in terms of VOC signature and JA profile but also in terms of their precursors synthesized through the lipoxygenase (LOX)-pathway at the early stage of the plant response. For this purpose, two leading pests of potato with distinct feeding behaviours were used: the Colorado Potato Beetle (Leptinotarsa decemlineata Say), a chewing herbivore, and the Green Peach Aphid (Myzus persicae Sulzer), a piercing-sucking insect. The volatile signatures identified clearly differ in function with the feeding behaviour of the attacker and the aphid, which causes the smaller damages, triggers the emission of a higher number of volatiles. In addition, 9-LOX products, which are usually associated with defence against pathogens, were exclusively activated by aphid attack. Furthermore, a correlation between volatiles and JA accumulation and the evolution of their precursors was determined. Finally, the role of the insect itself on the plant response after insect infestation was highlighted.

Potato (Solanum tuberosum L.) tuber ageing induces changes in the proteome and antioxidants associated with the sprouting pattern.

During post-harvest storage, potato tubers age as they undergo an evolution of their physiological state influencing their sprouting pattern. In the present study, physiological and biochemical approaches were combined to provide new insights on potato (Solanum tuberosum L. cv. Désirée) tuber ageing. An increase in the physiological age index (PAI) value from 0.14 to 0.83 occurred during storage at 4 degrees C over 270 d. Using this reference frame, a proteomic approach was followed based on two-dimensional electrophoresis. In the experimental conditions of this study, a marked proteolysis of patatin occurred after the PAI reached a value of 0.6. In parallel, several glycolytic enzymes were up-regulated and cellular components influencing protein conformation and the response to stress were altered. The equilibrium between the 20S and 26S forms of the proteasome was modified, the 20S form that recycles oxidized proteins being up-regulated. Two proteins belonging to the cytoskeleton were also differentially expressed during ageing. As most of these changes are also observed in an oxidative stress context, an approach focused on antioxidant compounds and enzymes as well as oxidative damage on polyunsaturated fatty acids and proteins was conducted. All the changes observed during ageing seemed to allow the potato tubers to maintain their radical scavenging activity until the end of the storage period as no accumulation of oxidative damage was observed. These data are interpreted considering the impact of reactive oxygen species on the development and the behaviour of other plant systems undergoing ageing or senescence processes.

Oxylipin profile and antioxidant status of potato tubers during extended storage at room temperature.

Potato tubers (cv. Bintje) (Solanum tuberosum L.) were stored under extreme conditions at 20 degrees C for 350 days without sprout inhibitors in order to assess whether aging- and/or senescence-related processes occurred. Under these extreme storage conditions, multiple sprouting followed by the formation of daughter tubers occurs. At the same time, an increase in respiration intensity, as evidenced by cytochrome c oxidase activity (E.C. 1.9.3.1), is observed, leading to a potential increase in reactive oxygen species (ROS) production. As polyunsaturated fatty acids are priority targets of oxidative attacks, the damage to lipids was assessed by oxylipin profiling in both free and esterified forms. Oxylipin profiling showed a predominance of linoleic acid-derived oxylipins and of 9-hydroxy and 9-hydroperoxy fatty acids in both free and esterified forms. No significant accumulation of individual oxylipin was observed 350 days after harvest. To further understand the absence of lipid breakdown products accumulation, the main enzymatic and non-enzymatic antioxidants were assessed. Antioxidant enzyme activities [superoxide dismutase (E.C. 1.15.1.1), catalase (E.C. 1.11.1.6.), ascorbate peroxidase (E.C. 1.11.1.11)] were enhanced during the advanced phase of aging. The main non-enzymatic antioxidant compound, ascorbate, decreased markedly in the early stages of storage, followed by a slower decline. Total radical scavenging activity was also maintained at the end of the storage period. Our results indicate that the enhanced aging process occurring during storage at room temperature does not seem to be associated with the changes classically encountered during leaf senescence or seed aging and that the observed degenerative processes do not surpass the protective potential of the tubers.

Changes in oxylipin synthesis after Phytophthora infestans infection of potato leaves do not correlate with resistance.

Oxylipins constitute a class of molecules notably involved in host-pathogen interactions. In the potato-Phytophthora infestans (Mont.) De Barry (P. infestans) relationships, the role of colneleic and colnelenic acids, two oxylipins resulting from the consecutive action of lipoxygenase (EC 1.13.11.12) and divinyl ether synthase (EC 1.-) on respectively linoleic and linolenic acids have been previously reported. In the present paper, five potato cultivars with contrasting resistance to P. infestans were submitted to infection. Lipoxygenase pathway response was studied at both transcriptional and metabolic levels. A Northern blot preliminary study revealed that lipoxygenase (lox1 and lox3) and divinyl ether synthase genes were clearly up-regulated 96h after leaf inoculation with P. infestans. Profiling of free and esterified oxylipins performed 24h, 48h, 72h and 96h after inoculation, showed that esterified oxylipins are mainly produced with 9-derivatives in higher concentrations (esterified forms of colnelenic acid, 9-hydroxy octadecatrienoic acid, 9-hydroperoxy octadecatrienoic acid). Oxylipin accumulation is undetectable 24h after infection, slightly detectable after 48h, reaching highest concentrations after 96h. Cultivars show slightly different oxylipin profiles but the concentration of individual oxylipins differs markedly 96h after infection. No correlation was found between P. infestans resistance levels and oxylipin synthesis rates or concentration. To assess local and systemic effects of colneleic acid application before P. infestans infection, Bintje cultivar was sprayed with colneleic acid 72h before inoculation. Both application modes (local and systemic) resulted in lipoxygenase pathway activation without affecting the resistance level to the pathogen.

Interaction between the barley allelochemical compounds gramine and hordenine and artificial lipid bilayers mimicking the plant plasma membrane.

Some plants affect the development of neighbouring plants by releasing secondary metabolites into their environment. This phenomenon is known as allelopathy and is a potential tool for weed management within the framework of sustainable agriculture. While many studies have investigated the mode of action of various allelochemicals (molecules emitted by allelopathic plants), little attention has been paid to their initial contact with the plant plasma membrane (PPM). In this paper, this key step is explored for two alkaloids, gramine and hordenine, that are allelochemicals from barley. Using in vitro bioassays, we first showed that gramine has a greater toxicity than hordenine towards a weed commonly found in northern countries (Matricaria recutita L.). Then, isothermal titration calorimetry was used to show that these alkaloids spontaneously interact with lipid bilayers that mimic the PPM. The greater impact of gramine on the thermotropic behaviour of lipids compared to hordenine was established by means of infrared spectroscopy. Finally, the molecular mechanisms of these interactions were explored with molecular dynamics simulations. The good correlation between phytotoxicity and the ability to disturb lipid bilayers is discussed. In this study, biophysical tools were used for the first time to investigate the interactions of allelochemicals with artificial PPM.

Decrease in the Photosynthetic Performance of Temperate Grassland Species Does Not Lead to a Decline in the Gross Primary Production of the Ecosystem.

Plants, under stressful conditions, can proceed to photosynthetic adjustments in order to acclimatize and alleviate the detrimental impacts on the photosynthetic apparatus. However, it is currently unclear how adjustment of photosynthetic processes under environmental constraints by plants influences CO2 gas exchange at the ecosystem-scale. Over a 2-year period, photosynthetic performance of a temperate grassland ecosystem was characterized by conducting frequent chlorophyll fluorescence (ChlF) measurements on three primary grassland species (Lolium perenne L., Taraxacum sp., and Trifolium repens L.). Ecosystem photosynthetic performance was estimated from measurements performed on the three dominant grassland species weighed based on their relative abundance. In addition, monitoring CO2 fluxes was performed by eddy covariance. The highest decrease in photosynthetic performance was detected in summer, when environmental constraints were combined. Dicot species (Taraxacum sp. and T. repens) presented the strongest capacity to up-regulate PSI and exhibited the highest electron transport efficiency under stressful environmental conditions compared with L. perenne. The decline in ecosystem photosynthetic performance did not lead to a reduction in gross primary productivity, likely because increased light energy was available under these conditions. The carbon amounts fixed at light saturation were not influenced by alterations in photosynthetic processes, suggesting photosynthesis was not impaired. Decreased photosynthetic performance was associated with high respiration flux, but both were influenced by temperature. Our study revealed variation in photosynthetic performance of a grassland ecosystem responded to environmental constraints, but alterations in photosynthetic processes appeared to exhibit a negligible influence on ecosystem CO2 fluxes.

Study of precursors responsible for off-flavor formation during storage of potato flakes.

Off-flavors frequently appear during the storage of potato flakes. Volatile profile analysis performed by solid-phase microextraction-gas chromatography-mass spectrometry revealed that hexanal is the main compound that appears during the storage period. Hexanal may be a degradation product of linoleic acid formed through linoleic acid hydroperoxide cleavage. Profiles of hexanal precursors were determined from potato flakes at different storage time points. Linoleic acid-derived oxylipins are predominant in potato flakes. The free oxylipins identified, in descending order, are as follows: hexanal, hydroxy polyunsaturated fatty (PUFAs), oxo PUFAs, divinyl ether PUFAs, and hydroperoxy PUFAs. However, the main oxylipins detected were esterified: esterified hydroxy, hydroperoxy, and oxo PUFAs. Oxylipins reveal different evolutions during the storage period. Chiral high-performance liquid chromatography analysis of the precursors of hexanal and other oxylipins revealed a racemic composition that supports the nonenzymatic formation of hexanal and most of the other oxylipins identified.

Barley (Hordeum distichon L.) roots synthesise volatile aldehydes with a strong age-dependent pattern and release (E)-non-2-enal and (E,Z)-nona-2,6-dienal after mechanical injury.

In the context of chemical ecology, the analysis of the temporal production pattern of volatile organic compounds (VOCs) in root tissues and the emission rate measurement of root-emitted VOCs are of major importance for setting up experiments to study the implication of these compounds in biotic interactions. Such analyses, however, remain challenging because of the belowground location of plant root systems. In this context, this study describes the evolution of the root VOC production pattern of barley (Hordeum distichon L.) at five developmental stages from germination to the end of tillering and evaluates the emission of the identified VOCs in an artificial soil. VOCs produced by crushed root tissues and released by unexcavated root systems were analysed using dynamic sampling devices coupled to a gas chromatography-mass spectrometry methodology (synchronous SCAN/SIM). The results showed that, at each analysed developmental stage, crushed barley roots produced mainly four volatile aldehydes: hexanal; (E)-hex-2-enal; (E)-non-2-enal; and (E,Z)-nona-2,6-dienal. Higher total and individual VOC concentrations were measured in 3-day-old seminal roots compared with older phenological stages. For each developmental stage, the lipoxygenase (LOX) activity was greater for linoleic acid than α-linolenic acid and the greatest LOX activities using linoleic and α-linolenic acids as substrates were measured in 7- and 3-day-old roots, respectively. The analysis of VOCs released by barley roots into the soil showed that (E)-non-2-enal and (E,Z)-nona-2,6-dienal were the only VOCs emitted in quantifiable amounts by mechanically injured roots.

Biotechnological uses of RNAi in plants: risk assessment considerations.

RNAi offers opportunities to generate new traits in genetically modified (GM) plants. Instead of expressing novel proteins, RNAi-based GM plants reduce target gene expression. Silencing of off-target genes may trigger unintended effects, and identifying these genes would facilitate risk assessment. However, using bioinformatics alone is not reliable, due to the lack of genomic data and insufficient knowledge of mechanisms governing mRNA-small (s)RNA interactions.

Morpho-histology and genotype dependence of in vitro morphogenesis in mature embryo cultures of wheat.

Cellular totipotency is one of the basic principles of plant biotechnology. Currently, the success of the procedure used to produce transgenic plants is directly proportional to the successful insertion of foreign DNA into the genome of suitable target tissue/cells that are able to regenerate plants. The mature embryo (ME) is increasingly recognized as a valuable explant for developing regenerable cell lines in wheat biotechnology. We have previously developed a regeneration procedure based on fragmented ME in vitro culture. Before we can use this regeneration system as a model for molecular studies of the morphogenic pathway induced in vitro and investigate the functional links between regenerative capacity and transformation receptiveness, some questions need to be answered. Plant regeneration from cultured tissues is genetically controlled. Factors such as age/degree of differentiation and physiological conditions affect the response of explants to culture conditions. Plant regeneration in culture can be achieved through embryogenesis or organogenesis. In this paper, the suitability of ME tissues for tissue culture and the chronological series of morphological data observed at the macroscopic level are documented. Genetic variability at each step of the regeneration process was evaluated through a varietal comparison of several elite wheat cultivars. A detailed histological analysis of the chronological sequence of morphological events during ontogeny was conducted. Compared with cultures of immature zygotic embryos, we found that the embryogenic pathway occurs slightly earlier and is of a different origin in our model. Cytological, physiological, and some biochemical aspects of somatic embryo formation in wheat ME culture are discussed.

Possible consequences of the overlap between the CaMV 35S promoter regions in plant transformation vectors used and the viral gene VI in transgenic plants.

Multiple variants of the Cauliflower mosaic virus 35S promoter (P35S) are used to drive the expression of transgenes in genetically modified plants, for both research purposes and commercial applications. The genetic organization of the densely packed genome of this virus results in sequence overlap between P35S and viral gene VI, encoding the multifunctional P6 protein. The present paper investigates whether introduction of P35S variants by genetic transformation is likely to result in the expression of functional domains of the P6 protein and in potential impacts in transgenic plants. A bioinformatic analysis was performed to assess the safety for human and animal health of putative translation products of gene VI overlapping P35S. No relevant similarity was identified between the putative peptides and known allergens and toxins, using different databases. From a literature study it became clear that long variants of the P35S do contain an open reading frame, when expressed, might result in unintended phenotypic changes. A flowchart is proposed to evaluate possible unintended effects in plant transformants, based on the DNA sequence actually introduced and on the plant phenotype, taking into account the known effects of ectopically expressed P6 domains in model plants.