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Polyphenols are plant secondary metabolites that function mostly as a general stress-induced protective mechanism. Polyphenols have also gained interest due to their beneficial properties for human health. Strawberry leaves represent an agro-industrial waste material with relevant bioactive polyphenol content, which could be incorporated into circular economy strategies. However, due to the low quantities of polyphenols in plants, their production needs to be improved for cost-effective applications. The objective of this research was to compare polyphenol production in strawberry (Fragaria × ananassa cv. Festival) leaves in plants grown in greenhouse conditions and plants grown in vitro, using three possible elicitor treatments (UV irradiation, cold exposure, and cysteine). General vegetative effects were morphologically evaluated, and specific polyphenolic compounds were quantified by UHPLC-DAD-MS/MS. Gallic acid was the most abundant polyphenol found in the leaves, both in vivo and in vitro. The results showed higher amounts and faster accumulation of polyphenols in the in vitro regenerated plants, highlighting the relevance of in vitro tissue culture strategies for producing compounds such as polyphenols in this species and cultivar.

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Despite the positive results of using elicitors to induce resistance against plant diseases, some factors have inhibited the popularization of their use in agriculture. There is an energetic cost related to the elicitors' induced response which can cause undesired effects on growth under low-pressure disease conditions. Elicitors can create phytotoxicity and show high variation in their efficiency between different genotypes within the same crop; in addition, the positive results related to the induced resistance may not repeat in field treatments, adding to the possibility that they are not economically viable. Thus, we carried out two experiments to investigate the technical and economic efficiency of acibenzolar-S-methyl (ASM) and its association with fungicides in the control of leaf diseases of susceptible and resistant wheat varieties, and as how it reflects on the photosynthetic and production performance of wheat. This study showed the limitations of incorporating ASM into foliar fungal disease control in economic terms. However, it was evident that ASM effectively induced plant resistance against Leaf Rust and Powdery Mildew in the field and can be considered a sustainable option for wheat cultivation. Even though its association with chemical control was not the best economic strategy the use of ASM is a tool that can be incorporated into wheat cultivation to minimize the emergence of fungicide-resistant pathogens due to the diversification of modes of action employed and reduce the toxic residue deposition to the environment and human health.

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Acmella radicans (Asteraceae) is a plant native to America. Despite it having medicinal attributes, studies on its phytochemical properties are scarce, and biotechnological studies do not exist for this species. In this study, we established an adventitious root culture from A. radicans internodal segments in shake flasks with indole-3-butyric acid (IBA), and then elicited it with jasmonic acid (JA) and salicylic acid (SA). The total phenolic content and antioxidant activity were evaluated, and a comparison was made using in vitro plantlets and wild plants. Internodal segments with 0.1 mg/L IBA showed 100% root induction and exhibited better growth after transfer to shake flasks with MS liquid culture medium. JA had a significant effect on biomass increase compared to unelicited roots, mainly with 50 µM JA (28%), while SA did not show significant results. Root elicited with 100 µM (SA and JA) showed a 0.34- and 3.9-fold increase, respectively, in total phenolic content (TPC) compared to the control. The antioxidant activity was also significant, and a lower half-maximal inhibitory concentration (IC50) was observed as the AJ concentration increased. Roots elicited with AJ (100 µM) exhibited high antioxidant activity with DPPH (IC50 = 9.4 µg/mL) and ABTS (IC50 = 3.3 µg/mL) assays; these values were close to those for vitamin C (IC50 = 2.0 µg/mL). The TPC and antioxidant activity of in vitro plants and root cultured in shake flasks showed the lowest values in most cases; even the root cultures without elicitation were better than those of a wild plant. In this study, we demonstrated that A. radicans root culture is capable of producing secondary metabolites, while its production and antioxidant activity can be enhanced using jasmonic acid.

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Public health, production and preservation of food, development of environmentally friendly (cosmeto-)textiles and plastics, synthesis processes using green technology, and improvement of water quality, among other domains, can be controlled with the help of chitosan. It has been demonstrated that this biopolymer exhibits advantageous properties, such as biocompatibility, biodegradability, antimicrobial effect, mucoadhesive properties, film-forming capacity, elicitor of plant defenses, coagulant-flocculant ability, synergistic effect and adjuvant along with other substances and materials. In part, its versatility is attributed to the presence of ionizable and reactive primary amino groups that provide strong chemical interactions with small inorganic and organic substances, macromolecules, ions, and cell membranes/walls. Hence, chitosan has been used either to create new materials or to modify the properties of conventional materials applied on an industrial scale. Considering the relevance of strategic topics around the world, this review integrates recent studies and key background information constructed by different researchers designing chitosan-based materials with potential applications in the aforementioned concerns.

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O objetivo neste trabalho foi verificar a influência da aplicação de diferentes concentrações de ácido salicílico e distintas coberturas de solo no cultivo de physalis (Physalis peruviana L). O trabalho foi desenvolvido na área experimental da Universidade Federal da Fronteira Sul, Laranjeiras do Sul-PR. Foram utilizadas mudas de physalis produzidas em casa de vegetação. A campo foram submetidas a cinco concentrações de ácido salicílico (0, 0,001, 0,002, 0,003, 0,004 Mol L-¹), e três coberturas de solo (palhada de gramínea, eucalipto e testemunha sem palhada). Nas avaliações de pré-colheita, o delineamento experimental foi de blocos ao acaso, em esquema fatorial 3x5. Para as avaliações pós-colheita o delineamento experimental foi 3x5x2, sendo considerado também o mês de colheita. Não houve interação entre os fatores para as variáveis analisadas. Para altura e diâmetro, presença de insetos, incidência e severidade de doenças foliares, somente o fator concentrações de ácido salicílico influenciou. Na pós-colheita somente o mês de colheita interferiu nos fatores, sendo a colheita de setembro com maior número e massa dos frutos, mas também o maior ataque de insetos e doenças. Concluiu-se com este trabalho que a aplicação de ácido salicílico influenciou no desenvolvimento de plantas de physalis.(AU)

The objective of this work was to verify the influence of the application of different concentrations of salicylic acid and different soil coverings in the cultivation of physalis (Physalis peruvianaL). The work was developed in the experimentalarea of the Federal University of Fronteira Sul, Laranjeiras do Sul-PR. Physalis seedlings produced in a greenhouse were used. The field was subjected to five concentrations of salicylic acid (0, 0,001, 0,002, 0,003, 0,004 Mol L-¹), and three soil coverings (grass straw, eucalyptus and control without straw). In the pre-harvest evaluations, the experimental design was randomized blocks, in a 3x5 factorial scheme. For post-harvest evaluations, the experimental design was 3x5x2, and the month of harvest was also considered. There was no interaction between the factors for the analyzed variables. For height and diameter increase and presence of insects, incidence and severity of leaf diseases, only the factor of salicylic acid influences. In the post-harvestperiod, only the month of harvest interfered in the factors, with the September harvest having the largest number and mass of fruits, but also the greatest attack by insects and diseases. It was concluded with this work that the application of salicylic acid influenced the development of physalis plants.(AU)

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The objective of this review is to present a compilation of the application of various biostimulants in strawberry plants. Strawberry cultivation is of great importance worldwide, and, there is currently no review on this topic in the literature. Plant biostimulation consists of using or applying physical, chemical, or biological stimuli that trigger a response-called induction or elicitation-with a positive effect on crop growth, development, and quality. Biostimulation provides tolerance to biotic and abiotic stress, and more absorption and accumulation of nutrients, favoring the metabolism of the plants. The strawberry is a highly appreciated fruit for its high organoleptic and nutraceutical qualities since it is rich in phenolic compounds, vitamins, and minerals, in addition to being a product with high commercial value. This review aims to present an overview of the information on using different biostimulation techniques in strawberries. The information obtained from publications from 2000-2022 is organized according to the biostimulant's physical, chemical, or biological nature. The biochemical or physiological impact on plant productivity, yield, fruit quality, and postharvest life is described for each class of biostimulant. Information gaps are also pointed out, highlighting the topics in which more significant research effort is necessary.

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Common bean (Phaseolus vulgaris L.), soybean (Glycine max L.) and mung bean (Vigna radiata L. Wilczek) seedlings were treated with methyl jasmonate (MeJA); then, dose-response and time-course experiments were carried out. Isoflavonoid composition was evaluated by high performance liquid chromatography. As a result of MeJA induction, all leguminous plants increase the amount of isoflavonoids, at levels that depend on the concentration of the elicitor and the time after induction. However, the application of MeJA in concentrations higher than 2.22 mM showed deleterious effects on seedlings and strong decreases in the concentration of isoflavonoids. In addition, a series of compounds structurally related to MeJA, such as jasmonic acid, cis-jasmone, coronatine, and indanoyl derivatives, were evaluated as elicitors. The results show that coronatine and the indanoyl-amino acids conjugates displayed a significant elicitor effect of isoflavonoids in common bean (cvs. Cargamanto Mocho and Corpoica LAS 106) and soybean (cv. Soyica P-34) seedlings, even higher than that found with the recognized elicitors, benzo (1,2,3) thiadiazole-7-carbothioic acid S-methyl ester (acibenzolar S-methyl) and benzo-(1,2,3) thiadiazole-7-carbothioic acid (acibenzolar acid). Leguminous plants can be treated with jasmonates and indanoyl derivatives to increase levels of bioactive isoflavonoids and consequently improve biological and functional properties and resistance against pests.

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RESUMEN La utilización de elicitores es frecuente en la inducción de diferentes respuestas relacionadas con la resistencia sistémica de las plantas; no obstante, no se ha caracterizado el gasto metabólico que generan en el cultivo de papa variedad Diacol Capiro, que es la más utilizada para el procesamiento industrial en Colombia, debido a su calidad y alta producción. En esta investigación, se evaluó el efecto de cuatro elicitores en el crecimiento y desarrollo del cultivo de papa, en la vereda El Toval, del municipio de Tutazá, Boyacá. El diseño experimental fue en parcelas divididas, en donde la parcela principal fue la etapa fenológica, en la que se hizo la aplicación (E1; formación de tallos secundarios), época dos (E2; floración) y época tres (E3; aplicación tanto en formación de tallos secundarios como en floración) y la subparcela, con los elicitores aplicados (glucosamina, ácido γ-amino butírico, fosfito de potasio y ácido salicílico) y el testigo sin aplicación. Los datos fueron tomados a los 60, 80 y 100 días después de la siembra, teniendo como variables: altura de la planta, número de foliolos, clorofilas totales, eficiencia fotosintética, biomasa seca y producción. La glucosamina fue el tratamiento que obtuvo menores resultados en la mayoría de variables evaluadas con respecto al testigo; asimismo, los elicitores tienen efectos diferentes, dependiendo del estado fenológico del cultivo, especialmente, en la época de formación de tallos secundarios. El ácido salicílico (AS) presentó un comportamiento estadísticamente similar al del testigo, mientras que los demás elicitores aplicados, disminuyeron la producción de papa.

ABSTRACT Elicitors are frequently employed for the induction of different responses related to the systemic resistance of plants, however, it has not been fully characterized in the metabolic expenditure generated in the cultivation of the potato variety Diacol Capiro, the most planted for industrial processing in Colombia due to its quality and production. In this research, the effect of four elicitors on the growth and development of the potato crop was evaluated, in the municipality of Tutazá, Boyacá. The experimental design was divided plots, being the main plot the phenological stage in which the application was made (E1; formation of secondary stems), season two (E2; flowering) and season three (E3; application both in formation of secondary stems as in flowering) and the subplot, the applied elicitors (glucosamine, γ-amino butyric acid, potassium phosphite and salicylic acid) and the control without application. Data were taken 60, 80 and 100 days after sowing, having as variables: plant height, number of leaflets, total chlorophylls, photosynthetic efficiency, dry biomass and production. With the use of glucosamine the lowest results were obtained in most of the variables evaluated with respect to the control, likewise, elicitors had different effects depending on the phenological state of the crop, especially at the moment of secondary stem formation. Salicylic acid (AS) presented a statistically similar behavior to that of the control while the other applied elicitors decreased potato production.

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Many natural compounds can activate the plant immunity, and for this reason, they have attracted special interest in crop disease management. Previously, we isolated from strawberry leaves an ellagitannin (HeT), which elicits plant defense responses. In this research, we investigated bioactive compounds from field-collected strawberry leaves capable of inducing defense responses in Arabidopsis thaliana against a bacterial pathogen. Methanolic extracts of strawberry leaves sampled at different months were obtained and compared. The highest content of total soluble phenolic compounds was found in the methanolic extracts of leaves sampled in December (DME). The defense response induced in A. thaliana by DME was attributed to two ellagitannins, the HeT and galloyl-HHDP-glucose. Both compounds exhibited phytoprotective effects against Pseudomonas viridiflava and induced the expression of PDF1.2 and PR1 genes. These results provide an economic value to strawberry leaves, normally discarded at the end of the harvest stage of the crop, as a raw material for plant health enhancer bioinputs.

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Agricultural systems face several challenges in terms of meeting everyday-growing quantities and qualities of food requirements. However, the ecological and social trade-offs for increasing agricultural production are high, therefore, more sustainable agricultural practices are desired. Researchers are currently working on diverse sustainable techniques based mostly on natural mechanisms that plants have developed along with their evolution. Here, we discuss the potential agricultural application of extracellular DNA (eDNA), its multiple functioning mechanisms in plant metabolism, the importance of hormetic curves establishment, and as a challenge: the technical limitations of the industrial scale for this technology. We highlight the more viable natural mechanisms in which eDNA affects plant metabolism, acting as a damage/microbe-associated molecular pattern (DAMP, MAMP) or as a general plant biostimulant. Finally, we suggest a whole sustainable system, where DNA is extracted from organic sources by a simple methodology to fulfill the molecular characteristics needed to be applied in crop production systems, allowing the reduction in, or perhaps the total removal of, chemical pesticides, fertilizers, and insecticides application.

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Asian Soybean Rust (ASR), a disease caused by Phakopsora pachyrhizi, causing yield losses up to 90%. The control is based on the fungicides which may generate resistant fungi. The activation of the plant defense system, should help on ASR control. In this study, secondary metabolites of Pseudomonas aeruginosa LV strain were applied on spore germination and the expression of defense genes in infected soybean plants. The F4A fraction and the pure metabolites were used. In vitro, 10 µg mL-1 of F4A reduced spore germination by 54%, while 100 µg mL-1 completely inhibited. Overexpression of phenylalanine ammonia lyase (PAL), O-methyltransferase (OMT) and pathogenesis related protein-2 (PR-2; glucanases) defense-related genes were detected 24 and 72 h after soybean sprouts were sprayed with an organocopper antimicrobial compound (OAC). Under greenhouse conditions, the best control was observed in plants treated with 60 µg mL-1 of PCA, which reduced ASR severity and lesion frequency by 75% and 43%, respectively. Plants sprayed with 2 and 20 µg mL-1 of F4A also decreased severity (41%) and lesion frequency (32%). The significant reduction in spore germination ASR in plant suggested that the strain of these metabolites are effective against P. pachyrhizi, and they can be used for ASR control.

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Bacterial genome sequencing has revealed a vast number of novel biosynthetic gene clusters (BGC) with potential to produce bioactive natural products. However, the biosynthesis of secondary metabolites by bacteria is often silenced under laboratory conditions, limiting the controlled expression of natural products. Here we describe an integrated methodology for the construction and screening of an elicited and pre-fractionated library of marine bacteria. In this pilot study, chemical elicitors were evaluated to mimic the natural environment and to induce the expression of cryptic BGCs in deep-sea bacteria. By integrating high-resolution untargeted metabolomics with cheminformatics analyses, it was possible to visualize, mine, identify and map the chemical and biological space of the elicited bacterial metabolites. The results show that elicited bacterial metabolites correspond to ~45% of the compounds produced under laboratory conditions. In addition, the elicited chemical space is novel (~70% of the elicited compounds) or concentrated in the chemical space of drugs. Fractionation of the crude extracts further evidenced minor compounds (~90% of the collection) and the detection of biological activity. This pilot work pinpoints strategies for constructing and evaluating chemically diverse bacterial natural product libraries towards the identification of novel bacterial metabolites in natural product-based drug discovery pipelines.

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Abstract Thevetia peruviana is a medicinal plant that has valuable secondary terpenoid-type metabolites and phenolic compounds. Some flavonoid compounds of pharmaceutical interest stand out in the latter group. The concentration of these bioactive compounds in natural conditions is limited by environmental; therefore, it has been considered necessary to make in vitro plant cell suspension cultures that admit the use of elicitors to increase the content of active principles. Accordingly, in this study, for the optimization of flavonoid production in cell suspension culture of T. peruviana, different parameters related to elicitation with methyl jasmonate (MeJa), and salicylic acid (SA) were evaluated, at stirred flask scale. Firstly, 3 μM MeJa and 300 μM SA were added separately in cell cultures of T. peruviana, to assess their potential effects. Secondly, several experimental conditions were evaluated, for optimization purpose. In the first part, MeJa and SA increased the total flavonoid content, in 1.07 and 1.3 times, respectively, compared to the control culture; in the second part, total flavonoid content produced in MeJa mediated cell suspension cultures were 4.14 mg QE/g DW (milligrams of quercetin equivalent per gram of dry biomass) with: concentration 0.3 μM, addition time day 5 and harvest time 90 h. On the other hand, total flavonoid content produced in SA mediated cell suspension cultures were 3.75 mg QE/g DW with: concentration 100 μM, addition time day 0 and harvest time 96 h. Elicitation of cell suspension cultures of T. peruviana with MeJa and SA under their ideal parameter values increased flavonoid content.

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As a result of the capability of fungi to respond to culture conditions, we aimed to explore and compare the antibacterial activity and chemical diversity of two endophytic fungi isolated from Hyptis dilatata and cultured under different conditions by the addition of chemical elicitors, changes in the pH, and different incubation temperatures. Seventeen extracts were obtained from both Pestalotiopsis mangiferae (man-1 to man-17) and Pestalotiopsis microspora (mic-1 to mic-17) and were tested against a panel of pathogenic bacteria. Seven extracts from P. mangiferae and four extracts from P. microspora showed antibacterial activity; while some of these extracts displayed a high-level of selectivity and a broad-spectrum of activity, Pseudomonas aeruginosa was the most inhibited microorganism and was selected to determine the minimal inhibitory concentration (MIC). The MIC was determined for extracts man-6 (0.11 µg/mL) and mic-9 (0.56 µg/mL). Three active extracts obtained from P. mangiferae were analyzed by Liquid Chromatography-Electrospray Ionization-Quadrupole-Time of Flight-Mass Spectrometry (LC-ESI-Q-TOF-MS) to explore the chemical diversity and the variations in the composition. This allows us to propose structures for some of the determined molecular formulas, including the previously reported mangiferaelactone (1), an antibacterial compound.

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Bemisia tabaci Gennadius (Hemiptera: Aleyrodidae) biotype B is a key pest of Solanum lycopersicum L. (Solanaceae) throughout the world. In this study, we examined the induction of resistance on tomato plants treated with SA, BABA, and Trichoderma either individually or in combination against B. tabaci biotype B through the assessment of some biological and behavioral aspects of this insect pest. Also, to understand the mode of action of these inducers, we correlated and analyzed the biochemical basis of plant resistance, by measuring levels of polyphenol oxidase (PPO), peroxidase (POD), phenylalanine ammonia lyase (PAL), and phenolic content in leaves of treated tomato plants. The longest development time of whitefly immature stages was recorded for plants treated with root ß-aminobutyric acid application (RBABA) + root Trichoderma application (RT), root salicylic acid application (RSA) + RT, and RT. In a free-choice assay, B. tabaci adults showed a significantly lower preference for settling and oviposition in RBABA + RT, RSA + RT, and RT in comparison with control. In a no-choice assay, B. tabaci females laid significantly fewer eggs on treatments than those in control, with better results observed in RBABA + RT. Plants responded to different treatments and showed higher induction of PPO, POD, and PAL activities, besides the higher accumulation of phenols in RBABA + RT, RSA + RT, and RT treatments. These results suggest that RBABA + RT, RSA + RT, and RT could be utilized for the induction of effective plant defense against B. tabaci.

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Herbaspirillum rubrisubalbicans is the causal agent of red stripe disease (RSD) and mottle stripe disease of sorghum and sugarcane, respectively. In all, 63 genotypes of Sorghum bicolor were inoculated with H. rubrisubalbicans, with 59 showing RSD symptoms. Quantitative trait loci (QTL) analysis in a recombinant inbred line (RIL) population identified several QTL associated with variation in resistance to RSD. RNA sequencing analysis identified a number of genes whose transcript levels were differentially regulated during H. rubrisubalbicans infection. Among those genes that responded to H. rubrisubalbicans inoculation were many involved in plant-pathogen interactions such as leucine-rich repeat receptors, mitogen-activated protein kinase 1, calcium-binding proteins, transcriptional factors (ethylene-responsive element binding factor), and callose synthase. Pretreatment of sorghum leaves with the pathogen-associated molecular pattern (PAMP) molecules flg22 and chitooctaose provided protection against subsequent challenge with the pathogen, suggesting that PAMP-triggered immunity plays an important role in the sorghum immunity response. These data present baseline information for the use of the genetically tractable H. rubrisubalbicans-sorghum pathosystem for the study of innate immunity and disease resistance in this important grain and bioenergy crop. Information gained from the use of this system is likely to be informative for other monocots, including those more intractable for experimental study (e.g., sugarcane).

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Chili pepper (Capsicum annuum L.) production is affected by wilt and root rot, the most devastating disease caused by the pathogen complex of oomycete Phytophthora capsici Leon and the fungi Fusarium oxysporum Schlecht and Rhizoctonia solani Kühn, infecting roots, stems, leaves, and fruits. Fungicides are currently inefficient against this disease and have a high environmental impact. The use of elicitors is a sustainable alternative for inducing resistance to wilting and root rot. DNA fragments of an organism's own origin (conspecific or self-DNA) have shown the ability to inhibit growth and activate defense mechanisms in some plant species. In this investigation, the effect of the fragmented DNA mixture of Phytophthora capsici L., Fusarium oxysporum S., and Rhizoctonia solani K. on the protection against wilt and root rot of Capsicum annuum L. plants was evaluated. Changes in plant performance, phenolics, and flavonoids contents, as well as gene expression involved in the production of defense metabolites after the fragmented and unfragmented DNA mixture in three concentrations (20, 60, and 100 µg mL-1) in chili peppers, were studied. The results obtained showed a decrease in plant height in 60 and 100 µg mL-1 concentrations in absence of pathogens. Moreover, the treatment with fragmented DNA 100 µg mL-1 showed significant increase in the content of phenolic compounds and total flavonoids as well as gene expression associated to plant defense in comparison with control plants. Interestingly, foliar application of DNA fragments of the pathogen complex to a concentration of 100 µg mL-1 caused a 40% decrease in the mortality of infected plants with the pathogens at 30 days post-inoculation compared with control plants inoculated with the pathogen complex but not sprayed with DNA fragments. These results suggested a perspective for application of fragmented DNA of these pathogens at the agricultural level in crop protection strategies to cope with wilt and root rot in Capsicum.

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The galling insect manipulates the host plant tissue to its own benefit, building the gall structure where it spends during most of its life cycle. These specialist herbivore insects can induce and manipulate plant structure and metabolism throughout gall development and may affect plant volatile emission. Consequently, volatile emission from altered metabolism contribute to eavesdropping cueing. Eavesdropping can be part of adaptive strategies used by evolution for both galling insects and the entire-associated community in order to cue some interaction response. This is in contrast to some herbivores associated with delayed induced responses, altering plant metabolites during the short time while they feed. Due to the different lifestyles of the galling organism, which are associated with different plant tissues and organs (e.g leaves, flowers or fruits), a distinct diversity of organisms may eavesdrop on induced volatiles interacting with the galls. Furthermore, the eavesdropping cues may be defined according to the phenological coupling between galling organism and host plant, which results from the development of a gall structure. For instance, when plants release volatile-induced defenses after galling insects' activity, another interactor may perceive these volatiles and change its behavior and interactions with host plants and galls. Thus, natural enemies could be attracted by different volatiles emitted by the gall tissues. Considering the duration of the life cycle of the galling organism and the gall, the temporal extent of gall-induced volatiles may include more persistent volatile cues and eavesdropping effects than the volatiles induced by non-galling herbivores. Accordingly, from chemical ecology perspective we expect that galling herbivore-induced volatiles may exhibit robust effects on neighboring-plant interactions including those ones during different plant developmental or phenological periods. Information about multitrophic interactions between insects and plants supports the additional understanding of direct and indirect effects, and allows insight into new hypotheses.

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The present work analyses the chromatographic profile of the peels from fruits of different citrus cultivated in Colombia: sweet orange (Citrus sinensis [L.] Osbeck var. Valencia), mandarins (Citrus reticulata L. var. Arrayana and Oneco), Key lime (Citrus aurantifolia [Christ.] Swingle var. Pajarito), Mandarine lime (Citrus x limonia, a hybrid between Citrus reticulata and Citrus x limon) and Tahitian lime (C. latifolia Tanaka, syn. Persian lime). Coumarins, furanocoumarins and polymethoxylated flavones are the major compounds. Then, six coumarins were isolated and identified from fruits of Tahitian and Key lime corresponding to 5-geranyloxy-7-methoxycoumarin; 5,7-dimethoxycoumarin (syn. limettin); 5,8-dimethoxypsoralen (syn. isopimpinellin); 5-methoxypsoralen (syn. bergaptene); 5-geranoxypsoralen (syn. bergamottin) and 5-(2,3-dihydroxy-3-methylbutoxy) psoralen (syn. oxypeucedanin hydrate). Coumarins and furanocoumarins were quantified by liquid chromatography (HPLC-DAD). Results show that the prenylated compounds were present in high concentrations in Tahitian and Key lime but in very low amounts in mandarins and sweet orange. Subsequently, the antifungal activity (inhibition of mycelial growth and germination of spores) of the coumarins against the fungus causing the anthracnose, Colletotrichum sp. (isolated from aerial parts of Tahitian lime) was determined. The compounds limettin and bergaptene, as well as mixtures of them, showed significant inhibitory effect (radial growth and spore germination) when compared to the control. Finally, the effect of some recognized elicitors to induce the coumarin production in fruits of C. latifolia was evaluated. The results showed that the chemical profiles are dependent on the applied elicitor and the post-induction time. As a result of the induction, a high concentration of some coumarins and furanocoumarins was maintained in the course of time for the Tahitian lime. In conclusion, isolated coumarins could be involved in the defense mechanisms of C. latifolia, C. aurantifolia and C. limonia and their accumulation may be modulated by the application of elicitors.

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OBJECTIVES: The aim of this study was to develop a Trifolium pratense hairy root (HR) production protocol and select HR lines with high isoflavone yield following elicitor treatments. RESULTS: We obtained 13 independent HR lines, producing approximately three times more isoflavonoids than seedlings (3.3 mg/g dry weight) and in which 27 isoflavonoids were detected. Each HR line had its own isoflavonoid profile. These lines produced as major components daidzein, genistein, formononetin and biochanin A. Sucrose, salicylic acid (SA), yeast extract (YE) and flagellin 22 (flg22) were tested as elicitors. Using SA 140 mg/L, allowed the maximum isoflavonoid production in plantlets (11.9 mg/g dry weight) but reduced root growth, possibly as a result of its toxicity. The highest isoflavone production in HR (27.9 mg/g dry weight) was obtained using sucrose 60 g/L, for 3.5 days. CONCLUSION: This work reports the high production of various isoflavonoids with T. pratense elicited HR cultures.

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