A Fast and Precise Method To Identify Indolic Glucosinolates and Camalexin in Plants by Combining Mass Spectrometric and Biological Information

In this manuscript, a fast and accurate identification and quantitation by mass spectrometry of indolic glucosinolates and camalexin involved in defense in Arabidopsis thaliana are described. Two elicitation systems, inoculation with Botrytis cinerea and treatment with AgNO3, were used in Col-0 wild-type and mutant genotypes impaired in the biosynthesis of the selected metabolites. Identification of analytes was carried out by nontargeted LC/ESI-QTOF-MS profiling. Confirmation of indolic glucosinolates and camalexin was achieved by their absence in the cyp79B2/B3 and pad3 mutants as well as their respective fragmentation upon collision-induced dissociation. Camalexin accumulation was induced only after AgNO3 treatment, whereas all indolic glucosinolates were constitutively present. Inoculation with Botrytis did not influence camalexin concentration but caused most aliphatic and indolic glucosinolates contents to decrease. Only the pen 3.1 mutant showed increased indolic glucosinolate levels after Botrytis or AgNO3 treatments. In addition, profiles of secondary metabolite in nontreated Col-0 and mutant plants were analyzed by means of partial least squares coupled to discriminant analysis (PLS-DA), and differences in the basal levels of indolic glucosinolates and tryptophan between cyp79B2/B3 plants and the rest of genotypes, including Col-0, were found. This probably has to be taken into consideration when comparing stress responses of Col-0 and cyp79B2/B3. The use of mutants carrying alterations in biosynthetic pathways is proposed as a useful strategy to identify secondary metabolites

Metabolomics of Disease Resistance in Crops

Plants are continuously exposed to the attack of invasive microorganisms, such as fungi or bacteria, and also viruses. To fight these attackers, plants develop different metabolic and genetic responses whose final outcome is the production of toxic compounds that kill the pathogen or deter its growth or semiotic molecules that alert other individuals of the same plant species. These molecules are derived from the secondary metabolism and their production is induced upon detection of a pathogen-associated molecular pattern (PAMP). These PAMPs are different molecules that are perceived by the host cell triggering defense responses. PAMP-elicited compounds are highly diverse and specific of every plant species and can be divided into preformed metabolites or phytoanticipins that are converted into toxic molecules upon pathogen perception, and toxic metabolites or phytoalexins that are produced only upon pathogen attack. Moreover, plant volatile emissions are also modified in response to pathogen attack to alert neighboring individuals or to make plants less attractive to pathogen vector arthropods. Plant metabolite profiling techniques have allowed the identification of novel antimicrobial molecules that are induced upon elicitation. However, more studies are required to assess the specific function of metabolites or metabolite blends on plant-microbe interactions

Metabolic and Regulatory Responses in Citrus rootstock in Response to Adverse Environmental Conditions

In response to adverse environmental conditions, plants modify their metabolism to adapt to the new conditions. To differentiate common responses to abiotic stress from specific adaptation to a certain stress condition, two citrus rootstocks (Carrizo citrange and Cleopatra mandarin) with a different ability to tolerate stress were subjected to soil flooding and drought, two water stress conditions. In response to these conditions, both genotypes showed altered root proline and phenylpropanoid levels, especially cinnamic acid, which was a common feature to Carrizo and Cleopatra. This was correlated with alterations in the levels of phenylpropanoid derivatives likely involved in lignin biosynthesis. In the regulatory part, levels of both stress hormones abscisic acid (ABA) and jasmonic acid (JA) decreased in response to soil flooding irrespective of the genotype’s relative flooding tolerance, but, on the other hand, the concentration of both metabolites increased in response to drought, showing a transient accumulation of JA after a few days and a progressive pattern of ABA increase. These responses are probably associated with different regulatory processes under soil flooding and drought. In addition, alterations in indole acetic acid (IAA) levels in citrus roots seemed to be associated with particular stress tolerance. Moreover, both genotypes exhibited a low degree of overlap in the metabolites induced under similar stress conditions, indicating a specific mechanism to cope with stress in plant species. Results also indicated a different metabolic basal status in both genotypes that could contribute to stress tolerance

Abscisic Acid as an Emerging Modulator of the Responses of Plants to Low Oxygen Conditions

Different environmental and developmental cues involve low oxygen conditions, particularly those associated to abiotic stress conditions. It is widely accepted that plant responses to low oxygen conditions are mainly regulated by ethylene (ET). However, interaction with other hormonal signaling pathways as gibberellins (GAs), auxin (IAA), or nitric oxide (NO) has been well-documented. In this network of interactions, abscisic acid (ABA) has always been present and regarded to as a negative regulator of the development of morphological adaptations to soil flooding: hyponastic growth, adventitious root emergence, or formation of secondary aerenchyma in different plant species. However, recent evidence points toward a positive role of this plant hormone on the modulation of plant responses to hypoxia and, more importantly, on the ability to recover during the post-hypoxic period. In this work, the involvement of ABA as an emerging regulator of plant responses to low oxygen conditions alone or in interaction with other hormones is reviewed and discussed

Teaching language and literature: A question of balance?

Regarding the teaching of Catalan, four textbooks were created in the 1970s and 80s, which included a complete work of literature, with numerous grammar and lexical commentaries, and exercises about the literary text. The author who is the link between these books is the grammarian Albert Jané i Riera, a writer and linguist who has been a leading figure in Catalan culture as the editor of the children's magazine Cavall Fort. Our aim with this synchronic and diachronic study is to vindicate the importance of the educational proposals we have analysed, as well as the appropriateness and the need of teaching language and literature to regain and consolidate their mutually enriching relationships

Educació literària i competència gramatical: una proposta d'ensenyament del català als anys setanta

Literature has often been one of the most relevant textual referents when teaching grammar. Even though teachers have often used literary texts as examples of linguistic elements in the didactic units included in language books for both primary and secondary education, it is rare for a single literary text to become a language teaching book. Yet this is what happened with the novel Abans de l'alba by Lluís Ferran de Pol (Arenys de Mar, 1911- 1995). De Pol's fictional text was first published in 1954 and reedited in 1973 as part of a collection devoted to the teaching of Catalan edited by writer, critic and educator Joan Triadú. This second edition, published by Aymà-Spes, transformed the novel into a Catalan language teaching book.The reasons for this change were due to its excellent prose, a fact that made it most valuable for language learning. Writer and language scholar Albert Jané and Ferran de Pol worked in close collaboration and changed the literary text into a successful language learningmanual by adding exercises and grammatical and lexical explanations

Rapid and reproducible determination of active gibberellins in citrus tissues by UPLC/ESI-MS/MS

Phytohormone determination is crucial to explain the physiological mechanisms during growth and development. Therefore, rapid and precise methods are needed to achieve reproducible determination of phytohormones. Among many others, gibberellins (GAs) constitute a family of complex analytes as most of them share similar structure and chemical properties although only a few hold biological activity (namely GA1; GA3; GA4 and GA7). A method has been developed to extract GAs from plant tissues by mechanical disruption using ultrapure water as solvent and, in this way, ion suppression was reduced whereas sensitivity increased. Using this methodology, the four active GAs were separated and quantified by UPLC coupled to MS/MS using the isotope-labeled internal standards [2 H2]-GA1 and [2 H2]-GA4. To sum up, the new method provides a fast and reproducible protocol to determine bioactive GAs at low concentrations, using minimal amounts of sample and reducing the use of organic solvents.Ministerio de Economia (MINECO) AGL2013-42038-R Universitat Jaume I P1IB2013-23 "Santiago Grisolia" grant from Generalitat Valenciana "Ramon y Cajal" contract from MINEC

Biochemical and hormonal changes associated with root growth restriction under cadmium stress during maize (Zea mays L.) pre-emergence

Cadmium (Cd) pollution of agricultural soils is a growing global concern. Plant growth restriction is the main visible symptom of Cd toxicity, and this metal may be particularly harmful to the preformed, seminal root during the pre-emergence stage. In the present study, we focused on Cd phytotoxicity in seminal root growth, nutrient composition, redox status, and hormone homeostasis during the pre-emergence stage of maize (Zea mays L) plants, distinguishing between the root apex and the remaining root tissue. After 72 h of metal exposure (50 and 100 µM CdCl2), root length and biomass, as well as Ca, Fe, Mg, and Mn contents, were diminished. A redox imbalance was evidenced by changes in peroxidase activities and the ascorbate–dehydroascorbate ratio decreased in both root parts. There were fewer carbonylated proteins in both root fractions after exposure to 50 µM Cd, compared to 100 µM Cd, which was related to increased 20S proteasome activities. Cd incremented ABA, IAA, and SA contents, but drastically reduced the biologically active gibberellin GA4 and the conjugate jasmonoyl-isoleucine (JA-Ile). We demonstrated that the whole root tissue is involved in the maize response to Cd stress, which entails redox and hormonal rearrangements, probably directed to widen the plant defense lines at the expense of root growth

Grafting improves tolerance to combined drought and heat stresses by modifying metabolism in citrus scion

Mediterranean basin and other citrus-growing areas, such as Florida or California, are among the most vulnerable regions to the impacts of global warming. Therefore, citrus will be likely subjected to increasing periods of water scarcity combined with high temperatures that will impair plant growth, development and yield. In citrus industry, grafting is used to improve varietal characteristics, such as abiotic stress tolerance. Previous research showed that under drought and heat stress conditions, Carrizo citrange (Poncirus trifoliata × Citrus sinensis) is a better rootstock than Cleopatra mandarin (Citrus reshni) because it induces a higher antioxidant activity on the scion, reducing oxidative damage and increasing plant tolerance. Here, it is shown that metabolic reconfiguration, including changes in carbohydrate and amino acid fluxes, are key responses for plant acclimation to stress conditions. Moreover, the importance of the rootstock on scion metabolic and hormonal responses to drought and heat stress combination has been addressed by using reciprocal grafting between Carrizo and Cleopatra genotypes. Thus, Carrizo as a rootstock improves the metabolic and hormonal response of Cleopatra scions to the stress combination by inducing the accumulation of protective compounds such as raffinose, galactinol and salicylic acid. In turn, Cleopatra as a rootstock reduces levels of raffinose, galactinol, proline, phenylalanine and tryptophan in Carrizo scions, which impairs plant tolerance to the stress combination. Our findings show the effect of the rootstock on scion metabolic response to stress combination and remark the importance of the rootstock in citrus plants exposed to harsh environmental conditions.Funding for open access charge: CRUE-Universitat Jaume

Effects of cadmium on gas exchange and phytohormone contents in citrus

The effect of increased Cd2+ concentrations in the watering solution on citrus physiology was studied by using two citrus genotypes, Cleopatra mandarin and Carrizo citrange. Cadmium content in roots and leaves was tested together with measurements of leaf damage, gas exchange parameters, and hormonal contents. Citrus roots efficiently retained Cd2+ avoiding its translocation to the shoots and Cleopatra mandarin translocated less Cd2+ than Carrizo. With increasing Cd2+ concentration all gas exchange parameters were decreased more in Carrizo than in Cleopatra mandarin. Cd-induced increases in abscisic acid and salicylic acid contents were observed in leaves but not in roots of both genotype