Chapter

Evaluation of Phytochemical and Biochemical Patterns of Lemon Verbena (Lippia citriodora H.B.K.) at Different Temperatures

Authors:
  • Medicinal Plants Research Center, Institute of Medicinal Plants, ACECR, Karaj, Iran
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Abstract

Background and objectives: Lemon verbena (Lippia citriodora H.B.K.) from Verbenaceae family, as an aromatic and medicinal plant, has attracted interests for its valuable essential oil (EO). This study was conducted to evaluate the effect of various temperatures on phytochemical, biochemical, and allometric traits of lemon verbena leaves. Methods: The experiment was designed on the basis of randomized complete block design (RCBD) with treatments of 5, 10, 15, 20, and 25 °C, and three replications. Results: The results showed that the EO content, main components, and chemical classes, except for oxygenated sesquiterpenes were enhanced by increasing the temperature from 5 to 25 ° C, while pigments, total soluble solid, proline, and soluble proteins were conversely decreased by increasing temperature. The highest fraction of variance among these variables was observed in the neral, EO, polyphenols and anthocyanins, respectively. According to cluster analysis (CA), the effect of temperature on the content of EO, main components, and chemical classes were classified into three groups (A: 5 and 10 °C, B: 15 and 20 °C, and C: 25 °C). Also, dendrogram cluster analysis showed three temperature groups (A: 5 °C, B: 10 °C, and C: 15-25 °C) on the basis of biochemical traits. Conclusion: The present study showed that the content of oxygenated sesquiterpenes and antioxidant pigments in contrast to the amount of EO were severely increased by decreasing the environmental temperature. These results clarify the quality and economic value of this plant at the time of harvesting and environmental conditions for the pharmaceuticals, health, and food industries.

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Low temperature is considered to be a key environmental factor that limits broad application of bermudagrass (Cynodon dactylon). The phytohormone ethylene has been confirmed to be involved in plant response to cold stress. However, there is limited knowledge concerning the proteomic alterations of ethylene-regulated cold stress response in plant. To explore the possible molecular mechanism, a proteomic approach was performed by using iTRAQ (isobaric tags for relative and absolute quantitation) system. Bermudagrass leaves were treated with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC) or water (as control) at 4 °C for 24 hours. Among 3,990 quantified proteins, 201 differentially expressed proteins by ethylene and cold stress were identified. To better understand the possible mechanism involved in the ethylene-regulated cold response in bemudagrass, eleven groups of differentially expressed proteins were further analyzed. These proteins were mainly related to lipid stability, antioxidant enzymes and antioxidants, ribosome pathway, as well as protein synthesis and degradation. Therefore, the process of ethylene-regulated cold response maybe mainly related to lipid peroxidation, stress response and denfence and protein metabolism. Taken together, this study provides some novel insights into the molecular mechanisms of ethylene in bermudagrass responses to cold stress.
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Metabolic acclimation of plants to cold stress may be of great importance for their growth, survival and crop productivity. The accumulation carbohydrates associated with cold tolerance (CT), transcript levels for genes encoding related enzymes along with damage indices were comparatively studied in three genotypes of bread and durum wheats differing in sensitivity. Two (Norstar, bread wheat and Gerdish, durum wheat) were tolerant and the other, SRN (durum wheat), was susceptible to cold stress. During cold stress (–5 °C for 24 h), the contents of electrolyte leakage index (ELI) in Norstar and then Gerdish plants were lower than that of SRN plants, particularly in cold acclimated (CA) plants (4 °C for 14 days), confirming lethal temperature 50 (LT50) under field conditions. Increased carbohydrate abundances in the cases of sucrose, glucose, fructose, hexose phosphates, fructan, raffinose, arabinose resulted in different intensities of oxidative stress in bread (Norstar) plants compared to durum plants (SRN and Gerdish) plants as well as in CA plants compared to non-acclimated (NA) ones under cold, indicating metabolic/regulatory capacity along with a decrease in ELI content and enhanced defense activities. A significant decrease in these carbohydrates, particularly sucrose, under cold in NA plants showed an elevated level of cell damage (confirmed by ELI) compared to CA plants. On the other hand, an increase in hexose phosphates, particularly in NA plants, indicated sucrose degradation along with greater production of glucose and fructose compared to CA plants. Under such conditions, a significant increase in transcript levels of sucrose synthase and acidic invertase confirmed these results. Under cold, the high ABA-containing genotypes like Norstar and then Gerdish, which were obvious in CA plants, partly induced relative acclimation of cells for acquisition of CT compared to SRN. These results reveal an important role of carbohydrate metabolism in creating CT in durum wheats (particularly in Gerdish) as well as bread wheat with possible responsive components in metabolic and transcript levels.
Article
Age-related chronic diseases associated to inflammation are becoming an important concern in modern societies due to the ageing of the population. Anoxybiotics (Anoxymins®) are a new class of bifunctional natural health ingredients, which are water-soluble carbohydrates from plants with antioxidative activity and additionally show anti-inflammatory and/or immunomodulatory effects. Anoxybiotics reduce oxidative distress, and have cell protective effects, which together with their additional biological activity (antiinflammation, immunomodulation, angiogenesis inhibition) result in a definite health benefit for the consumer. Planox L is a water-soluble extract from Lippia citriodora standardized in 10% Anoxyminis® developed by Anoxymer in collaboration with NDN Nutraceuticals and the Institute of Molecular and Cellular Biology. Planox L and its main bioactive oligosaccharide Anox-1 have demonstrated to be antiinflammatory (CAM-assay), antioxidant and joint-protecting through several in vitro and in vivo bioassays. The biological performance of Planox L has been also supported by clinical trials. In addition, toxicological tests have revealed the complete absence of toxicity. The application fields of this extract are cosmetics (creams, shampoo and sun milk) and functional food for joint-health (bakery, beverages and dairy) as antioxidant, anti-inflammation and anti-ageing agent. Particularly, due to their antioxidant/ antiinflammatory properties, Anoxymins® and therefore Planox L are focused on joint health (elderly and sports people).
Article
Purpose – The purpose of this paper is to determine the total phenol content (TP) and total flavonoid content (TF) fluctuations in different infusions of lemon verbena. Dried leaves were harvested in May, July and October and prepared at various soaking times (5-20 min). Design/methodology/approach – The Folin-Ciocalteu method was used to determine TP, while TF was determined by the aluminum chloride method. Findings – According to the results, the July harvest had the highest content of TP and TF, followed by May and October. Temperature and soaking time play a role in the proportion of released phenols and flavonoids (p < 0.05), whereas the content in continuous infusion is higher than infusion (without boiling) .TP and TF in infusion increase until 20 min, while concentration is convergent in continuous infusion after10 minutes, where no significant difference is seen. In the flowering stage, TP is close in comparison with the vegetative stage. Originality/value – Harvest time plays a role in the content, so it is best to prepare infusion from lemon verbena before the flowering period, when people drink it, as the results show. According to the authors’ knowledge, this is the first work that studied the variation in phenol and flavonoid content during different stages of growth with soaking time in drenched lemon verbena leaves.
Article
Pinus densiflora var. zhangwuensis grows fast, and its drought and salinity resistance are better than Pinus sylvestris var. mongolica. We compared cold hardiness and mechanisms of cold hardiness between the two species, to provide a theoretical basis for promoting and applying P. densiflora var. zhangwuensis in cold regions. A cold stress experiment was carried out on 3-year-old plantlets of P. densiflora var. zhangwuensis and P. sylvestris var. mongolica after hardening at five temperature regimes, 5, −10, −20, −40, and −60 °C, respectively. Some indices of needle samples for both species were measured, such as relative conductivity (REL), maximum photochemical efficiency (Fv/Fm), malondialdehyde (MDA), catalase (CAT), proline (Pro), soluble sugar (SS), and stomata density. REL and MDA values of both species after hardening had the same trend of increasing, but the trend was opposite in Fv/Fm value with increasing cold stress. Compared with P. sylvestris var. mongolica, the P. densiflora var. zhangwuensis had smaller increases in REL and MDA, and a smaller decline in Fv/Fm during cold stress. Compared to the control, REL growth of P. densiflora var. zhangwuensis and P. sylvestris var. mongolica at −60 °C were 0.41 and 0.60, and MDA growth was 29.94 mol g−1 FW and 47.80 mol g−1 FW, and Fv/Fm declines were 0.08 and 0.27. Half-lethal temperatures (LT50) calculated by logistic equation for P. densiflora var. zhangwuensis and P. sylvestris var. mongolica were −58.23 and −50.34 °C, respectively. These data suggest that cold resistance of P. densiflora var. zhangwuensis is stronger than that of P. sylvestris var. mongolica. Cold-resistance mechanisms of the two species differed. In response to cold stress, P. sylvestris var. mongolica had strong osmotic adjustment ability because of higher Pro and SS content, while P. densiflora var. zhangwuensis had strong antioxidant ability due to stronger CAT activity. Stomata density and diameter of P. densiflora var. zhangwuensis were smaller, as were single leaf area and number of leaves per plant, both characteristics promoting survival in a cold environment. Greater shoot height and total biomass of seedlings of P. densiflora var. zhangwuensis might be another reason for its stronger cold tolerance. © 2015, Northeast Forestry University and Springer-Verlag Berlin Heidelberg.
Article
Global climate change brings with it unwarranted shifts in both abiotic (heat stress, cold stress, wind, precipitation) and biotic (pathogens, pests) environmental factors, thus posing a threat to agricultural productivity across the world. In plants, lodging due to storms or herbivory causes wounding stress and consequently enhances endogenous jasmonates. In response, the plant growth is arrested as plant defense is prioritized. We pre-treated pea plants with elevated methyl jasmonate (MeJA) levels i.e. 50 μM, 100 μM and 200 μM under controlled growth chamber conditions. The pre-treated plants were then kept at 40 °C (heat stress-HS), 4 °C (cold stress-CS) and 20 °C (optimum/control temperature-OT) for 72 h. The effect of such treatments on plant growth attributes, photosynthesis, stomatal conductance, cell death rate, and regulation of endogenous hormones were observed. Elevated MeJA application hindered plant growth attributes under HS, CS and OT conditions. Moreover, elevated MeJA levels lowered the rate of photosynthesis and stomatal conductance, induced stomatal closure, caused higher cells mortality in leaves under HS, CS, and OT conditions. Endogenous ABA contents significantly declined in all MeJA treatments under HS and OT, but increased under CS conditions. Exogenous MeJA enhanced endogenous jasmonic acid contents of pea plants, but altered endogenous salicylic acid contents under varying temperatures. Current study shows that higher concentrations of exogenous MeJA strengthen plant defense mechanism by hindering plant growth under stress conditions.
Article
Acteoside (verbascoside) was isolated as an analgesic principle from Cedron (leaves and stem of Lippia triphylla (L'HER) O. KUNTZE; Verbenaceae), a Peruvian medicinal plant, by activity-guided separation. The compound exhibited analgesia on acetic acid-induced writhing and on tail pressure pain in mice by the oral administration of 300mg/kg and 100mg/kg, respectively. Acteoside also caused weak sedation by its effect on the prolongation of pentobarbital-induced anesthesia and on the depression of locomotion enhanced by methamphetamine. An intravenous injection of acteoside reduced the effective dose to 2mg/kg by the writhing method. Thirteen related compounds were tested for the activity by intravenous and oral administration to obtain information on the active structure.
Article
Low temperature is one of the most severe environmental factors that impair growth of plants and agricultural production. To investigate how actively growing birch (Betula platyphylla Suk) adapts to cold stress, two and a half-month-old birch plants were exposed to cold stress (4°C) and compared to plant material maintained at the control temperature (23°C). This treatment did not affect the survival of the plants, but growth was almost arrested. Relative electrolyte leakage (REL) of adult leaves after stress treatment of 4, 7, and 10 days at 4°C was significantly higher than REL of the leaves exposed to the control temperature, There was no significant difference, after 14 days, in REL of leaves exposed either to the control temperature or to cold stress at 4°C. These results show that birch can adapt to cold stress. Proteomic analysis, by bidimensional electrophoresis was performed, and a total of 15 protein spots were identified by mass spectrometry after 14 days of cold stress at 4°C. Proteins that were identified were involved in defense, photosynthesis, biosynthesis, carbon (C)-nitrogen (N) metabolism, and signal transduction. These proteins may be used for the establishment of a new network based on cooperation when plants are subjected to cold stress. Cold stress response proteins such as the β subunit of ATP synthase were mostly related to all aspects of chloroplast physiology, indicating that the cold resistance of birch was influenced, in part at least, by the chloroplast function. It was further found that the protein spots involved in defense responses, biosynthesis, C-N metabolism, and signal transduction were increased in intensity after cold stress, indicating that they played a key role in the cold hardiness mechanism of birch.
Article
The effects of shadecloth tree shelters on cold-induced photoinhibition, foliar anthocyanin and growth of Eucalyptus globulus Labill. and Eucalyptus nitens (Deane & Maiden) Maiden seedlings were assessed between planting (in early spring) and the age of 23 weeks. The experimental site was at 350 m above sea level (asl), which is considered marginal for establishment of E. globulus (but not E. nitens) plantations in Tasmania because of low mean minimum temperatures. Conditions within 3 weeks of planting induced severe photoinhibition in non-shaded seedlings. This was associated with increased anthocyanin and photodamage in non-shaded E. nitens and E. globulus. As a result, there was 20% mortality in non-shaded E. globulus. In contrast, shaded seedlings of both species had levels of photoinhibition and anthocyanin that were largely similar to those before planting and there was no photodamage. Levels of anthocyanin indicated that its synthesis responded to the severity of photoinhibition. Height growth and levels of mortality indicated that cold-induced photoinhibition, and not frost tolerance alone, determines the range of environments where E. globulus can be successfully planted. In contrast, the tolerance of E. nitens seedlings to cold-induced photoinhibition may be a factor in the demonstrated success of this species as a high-altitude plantation species.
Article
Long days (16 h light), high photon flux density (1200 µEm-2 s-1.) and high night temperature (20°C) resulted in the highest oil yield. : Daylength, night temperature, day temperature and photon flux density were important interacting factors determining oil composition. The photosynthate model proposed by Burbott and Loomis (Plant Physiol., 1967, <B.42, 20-8) explained the effect of environmental factors with respect to pulegone, menthone and menthofuran. Factors favouring the maintenance of high levels of photosynthate resulted in high concentrations of menthone and low concentrations of pulegone and menthofuran. The photosynthate model does not explain the effect of environmental factors on several other monoterpenes.
Article
Variations in oil content and the chemical composition of the oil of two chemotypes (G-1 and G-2) of scented geranium were investigated in different seasons in a semiarid tropical climate in south India. The results showed that weather parameters such as atmospheric temperatures and rainfall have influenced the oil content and its chemical composition. Increases in maximum temperatures reduced oil content in G-1, increased citronellol in G-1 and G-2, and increased citronellyl formate in G-1. Rainfall increased geraniol in G-2. Geraniol and citronellol in G-1 were highly negatively correlated.
Article
Exposure of plants to light intensities higher than those required to saturate photosynthesis leads to a reduction in photosynthetic capacity. This effect is known as photoinhibition. Photoinhibition is followed by destruction of carotenoids, bleaching of chlorophylls and increased lipid peroxidation due to damage by oxygen-derivatives. The oxygen concentration in chloroplasts in the light is high because of oxygen production by photosystem II (PSII). This can result in the release of reactive intermediates of reduced dioxygen such as superoxide radicals, hydroxyl radicals, hydrogen peroxide or singlet oxygen. In order to maintain their normal function under light stress conditions, chloroplasts have developed multiple repair and protection systems. The induction of specific light stress proteins, the ELIPs (for early light-induced proteins) can be considered to be part of these protective responses. The accumulation of ELIPs under light stress conditions is correlated with the photoinactivation of PSII, degradation of the Dl-protein of PSII reaction centre and changes in the level of pigments. Futhermore, the accumulation of ELIPs in the thylakoids is strictly controlled by the pigment content, especially by chlorophylls. Isolation of ELIPs in a native form and analysis of pigments bound to these proteins revealed that ELIPs can bind chlorophyll a and lutein. These data indicate that ELIPs might represent unique chlorophyll-binding proteins which have a transient function(s) during light stress. A transient ‘pigment-carrier’ function is postulated for ELIPs.
Article
In order to study the effects of irrigation levels on essential oil of balm (Melissa officinalis L), research was performed under field condition at Karaj, Iran in 2005. Experiment was conducted using randomized complete block design with 4 replications. Treatment included T1 (non stress), T2 (80%FC), T3 (60%FC), T4 (40%FC) and T5 (20%FC). Effects of irrigation levels on shoot yield, essential oil yield, essential oil percent, leaf yield, height plant, tiller number, stem diameter, stem yield and internode length was significant (á=1%). irrigation levels not significant effects on number of lateral stem number. Comparison of treatment means showed that highest shoot yield and height plant were relate to T1 (non stress), highest essential oil yield was achieved under T4 (40%FC) and essential oil percent related to T5 (20%FC). Highest stem diameter was related to T5 (20%FC). It could be concluded that moderate drought stress is beneficial for balm essential oil.
Article
Four inbred maize lines differing in chilling tolerance were used to study changes in water status and abscisic acid (ABA) levels before, during and after a chilling period. Seedlings were raised in fertilized soil at 24/22°C (day/night), 70% relative humidity. and a 12-h photoperiod with 200 μmol m−2 s−1 from fluorescent tubes. At an age of 2 weeks the plants were conditioned at 14/12°C for 4 days and then chilled for 5 days at 5/3°C. The other conditions (relative humidity, quantum flux, photoperiod) were unchanged. After the chilling period the plants were transferred to the original conditions for recovery. The third leaves were used to study changes in leaf necrosis, ion efflux, transpiration, water status and ABA accumulation. Pronounced differences in chilling tolerance between the 4 lines as estimated by necrotic leaf areas, ion efflux and whole plant survival were observed. Conditioning significantly increased tolerance against chilling at 5/3°C in all genotypes. The genotypes with low chilling tolerance had lower water and osmotic potentials than the more tolerant genotypes during a chilling period at 5/3°C. These differences were related to higher transpiration rates and lower diffusive resistance values of the more susceptible lines. During chilling stress at 5/3°C ABA levels were quadrupled. Only a small rise was measurable during conditioning at 14/12°C. However, conditioning enhanced the rise of ABA during subsequent chilling. ABA accumulation in the two lines with a higher chilling tolerance was triggered at a higher leaf water potential and reached higher levels than in the less tolerant lines. We conclude that chilling tolerance in maize is related to the ability for fast and pronounced formation of ABA as a protective agent against chilling injury.
Article
Five-day-old etiolated cucumber (Cucumis sativus L.) seedlings cv. Marketmore held at 2°C for 72 h developed chilling injury, resulting in desiccation and collapse of the hypocotyl tissue and eventual plant death. Hypoxia-induced accumulation of ethanol and acetaldehyde led to tolerance of subsequent chilling, as evidenced by continued hypocotyl growth and freedom from injury. Attenuated accumulation of volatiles by applied bisulfite reduced the development of hypoxia-induced chilling tolerance in seedlings. In seedlings held in normoxia cold tolerance was induced by applied ethanol vapors, whereas acetaldehyde had a marginal effect, suggesting that hypoxia-induced cold tolerance may arise from the accumulation and activity of ethanol. Cold tolerance was also induced by exposure of seedlings to volatile anesthetics including n-propanol, n-butanol, chloroform and halothane, suggesting that ethanol activity may result from fluidization of membrane lipids. This view is consistent with results which showed that ethanol activity was not associated with lipid metabolism. However, development of cold tolerance in ethanol-enriched tissues was time dependent, indicating that ethanol activity probably also entails biosynthetic event(s).
Article
This review provides a summary of the physiological dynamics andregulation of essential oil production, from the literature and availableinformation on diverse volatile oil crops. Essential oil production is highlyintegrated with the physiology of the whole plant and so depends on themetabolic state and preset developmental differentiation programme of thesynthesising tissue. Essential oil productivity is ecophysiologically andenvironmentally friendly. These and other aspects of the modulation ofessentialoil production are presented, along with a brief outline of the current conceptof the relevant biosynthetic mechanisms.
Article
Proline, which increases proportionately faster than other amino acids in plants under water stress, has been suggested as an evaluating parameter for irrigation scheduling and for selecting drought-resistant varieties. The necessity to analyze numerous samples from multiple replications of field grown materials prompted the development of a simple, rapid colorimetric determination of proline. The method detected proline in the 0.1 to 36.0 moles/g range of fresh weight leaf material.
Article
The losses of volatile constituents in herbs and spices depend mainly on drying parameters and biological characteristics of the plants. In the present study, two methods were applied in the analysis of the effect of drying on the aroma constituents of the widely used herbs thyme (Thymus vulgaris L.) and sage (Salvia officinalis L.). The volatile constituents of herbs (fresh, freeze-dried and ovendried at 30 °C and 60 °C) were isolated by dynamic headspace and simultaneous distillation-extraction methods and analysed by capillary gas chromatography and coupled gas chromatography-mass spectrometry. In total, 68 compounds were identified in thyme and 44 in sage, and more than 100 components were screened quantitatively. A significant reduction in the amount of extracted volatiles was found only in the case of drying at 60 °C, mainly as a result of the loss of non-oxygenated monoterpenes. The character of the changes of the headspace volatiles was more complex, especially for thyme, the content of aroma compounds being the highest when the herb was dried at 60 °C. Some aroma assessment criteria (coefficient of efficiency Ce) are proposed on the basis of the results obtained.
Article
The chemical composition of the essential oil extracted from fresh leaves of Lippia citriodora (Verbenaceae) was analyzed by GC-FID and GC–MS in May, when growth rates are maximal, and in September, in full bloom. In both samples the main constituents were geranial, neral and limonene constituting 66.3% of the total essential oil yield in May and increasing to 69% in September. Their individual percentage values, however, changed considerably for geranial and neral decreasing from 38.7 to 26.8% and from 24.5 to 21.8%, respectively, and for limonene increasing from 5.8 to 17.7%. All other components remained more or less unchanged both qualitatively and quantitatively. FT-IR spectrometry was also applied for the qualitative determination of the main components.
Article
This study selected 13 fruits and vegetables to determine their total phenolic and flavonoid contents and their stimulatory effects on splenocyte proliferation from female BALB/c mice. The highest total phenolic content was observed in mulberry (1515.9 ± 5.7 mg gallic acid equivalents (GAE)/100 g fresh matter (FM)) among four selected fruit species. The highest total phenolic content was observed in a variety of red onions (310.8 ± 4.9 mg GAE/100 g FM) among nine selected vegetable species. The highest total flavonoid content was observed in mulberry (250.1 ± 6.3 mg quercetin equivalents (QE)/100 g FM) among the selected fruits. The highest total flavonoid content was observed in ceylon spinach (133.1 ± 26.2 mg QE/100 g FM) among the selected vegetables. The mulberry, strawberry and red onion demonstrated an immuno-modulatory potential via stimulating splenocyte proliferation. Bitter melon showed a significantly (P < 0.05) negative correlation with splenocyte proliferation. Their immuno-modulatory components are highly correlated with phenolics, including flavonoids. The total phenolic contents in all selected fruits and vegetables significantly correlated with splenocyte proliferation in vitro.