Effects of low UV-B doses on the accumulation of UV-B absorbing compounds and total phenolics and carbohydrate metabolism in the peel of harvested lemons

This paper examined the peel (albedo and flavedo) of postharvest lemon fruits after UV-B exposure in order to analyze relationships between soluble carbohydrate metabolism and secondary metabolite accumulation. Lemons (Citrus limon, cv. Limoneira 8A) were harvested in winter months (June to August), treated with 0.43 W m⁻² (22 kJ m⁻² d⁻¹ UV-BBE) of UV-B radiation during 0 (control), 0.5, 1.0, 2.0, 3.0, and 5.0 min, and then stored at 25 °C for 24 h. Peel samples from irradiated areas were obtained with a razor blade and frozen in liquid nitrogen until use for measurements. Data obtained showed that 2 and 3 min of UV-B exposure effectively increased the level of UV-B absorbing compounds and total phenolics in flavedo without causing visual alterations of the peel colour as compared with non-irradiated lemons. By contrast, there were no significant changes in albedo secondary metabolite accumulation. The amount of secondary metabolites was depending upon UV-B time–dose. Exposure over 3.0 min did not further improve the accumulation of UV-B absorbing and phenolic compounds. Soluble sugars (sucrose, glucose and fructose) also accumulated in the lemon peel after UV-B exposure, but the distribution patterns were different. After 3 min time–dose, sucrose and hexoses increased in flavedo, whereas in albedo only increased the sucrose and glucose. This effect was related to UVB-induced changes in the activity of sucrose-hydrolyzing and sucrose-synthesizing enzymes: soluble and cell-bound invertase, sucrose synthase (SS) and sucrose phosphate synthase (SPS). Data indicate that lemon peel retains the capacity to modify the enzyme activity of sucrose metabolism in response to UV-B exposure. Our results also suggest that the exposure of postharvest lemons to low supplemental UV-B doses produces changes in the carbon allocation of peel tissues including synthesis, but probably not only limited to them, of UV-B absorbing and phenolic compounds.     

Usefulness of manufactured tomato extracts in the diagnosis of tomato sensitization: Comparison with the prick-prick method

Background

Commercial available skin prick test with fruits can be negative in sensitized or allergic patients due to a reduction in biological activity during the manufacturing process. Prick-prick tests with fresh foods are often preferred, but they are a non-standardized procedure. The usefulness of freeze-dried extracts of Canary Islands tomatoes, comparing the wheal sizes induced by prick test with the prick-prick method in the diagnosis of tomato sensitization has been analyzed. The objective of the study was to assess the potential diagnostic of freeze-dried extracts of Canary Islands tomatoes, comparing the wheal sizes induced by prick test with the prick-prick method.

Methods

Two groups of patients were analyzed: Group I: 26 individuals reporting clinical symptoms induced by tomato contact or ingestion. Group II: 71 control individuals with no symptoms induced by tomato: 12 of them were previously skin prick test positive to a tomato extract, 39 were atopic and 20 were non-atopic. All individuals underwent prick-prick with fresh ripe peel Canary tomatoes and skin prick tested with freeze-dried peel and pulp extracts obtained from peel and pulp of Canary tomatoes at 10 mg/ml. Wheal sizes and prick test positivity (≥ 7 mm²) were compared between groups.

Results

In group I, 21 (81%) out of 26 patients were prick-prick positive. Twenty patients (77%) had positive skin prick test to peel extracts and 12 (46%) to pulp extracts. Prick-prick induced a mean wheal size of 43.81 ± 40.19 mm² compared with 44.25 ± 36.68 mm² induced by the peel extract (Not significant), and 17.79 ± 9.39 mm² induced by the pulp extract (p < 0.01). In group II, 13 (18%) out of 71 control patients were prick-prick positive. Twelve patients (all of them previously positive to peel extract) had positive skin prick test to peel and 3 to pulp. Prick-prick induced a mean wheal size of 28.88 ± 13.12 mm² compared with 33.17 ± 17.55 mm² induced by peel extract (Not significant), and 13.33 ± 4.80 mm² induced by pulp extract (p < 0.05 with peel extract and prick-prick).

Conclusion

Canary peel tomato extract seems to be as efficient as prick-prick tests with ripe tomatoes to diagnose patients sensitized to tomato. The wheal sizes induced by prick-prick and peel extracts were very similar and showed a high correlation coefficient.     

Growth inhibitory effect of peel extract from Citrus junos

Extract from yuzu fruit peel (Citrus junos Sieb. ex Tanaka) strongly suppressed the germination of lettuce seeds while that from the peel of other citrus fruits such as navel orange (C. sinensis) and lemon (C. limon Burm. f.) had very little or no effect. The highest inhibitory activity was located in the peel followed by the segment but no significant activity was found in the seed extract of yuzu fruit. The effect of yuzu peel extract was tested on a variety of major crops including 38 crop species. Germination of seeds of tomato (Lycopersicon esculentum Mill.), celery (Apium graveolens L. var. dulce) and watercress (Nasturtium officinale R. Br.) was completely blocked by 27.8 mg dry peel equivalent ml^–1 of yuzu peel extract, while that of cucumber (Cucumis sativus L.) and pumpkin (Cucurbita moschata [Duch. ex Lam.] Duch. ex Poir.) seeds was not affected at this concentration, showing a broad variation in the sensitivity of crop seeds to the extract. The effectiveness of yuzu peel was also tested on some harmful weeds. The extract inhibited the elongation of both radicle and hypocotyl in summer weed quinoa (Chenopodium quinoa) grown on agar plates. Yuzu peel powder added to the soil was also effective in suppressing the growth of C. quinoa, Sonchus oleraceus L. and Digitaria ciliaris, while it had little effect on the spring weed Alopecurus aequalis Sobol. A strong growth inhibitory activity of yuzu peel was found in the aqueous phase after solvent extraction and most of the activity was recovered in the neutral fraction that passed through cation and anion exchange resins.     

Characterization of monoterpene biotransformation in two pseudomonads

Aims: To study the metabolic profile of Pseudomonas rhodesiae and Pseudomonas fluorescens in water–organic solvent systems using terpene substrates for both growth and biotransformation processes and to determine the aerobic or anaerobic status of these degradation pathways. Materials and Methods: Substrates from pinene (α‐pinene, α‐pinene oxide, β‐pinene, β‐pinene oxide, turpentine) and limonene (limonene, limonene‐1,2‐oxide, orange peel oil) families were tested. For the bioconversion, the terpene‐grown biomass was concentrated and used either as whole cells or as a crude enzymatic extract. Conclusion: Pseudomonas rhodesiae was the most suitable biocatalyst for the production of isonovalal from α‐pinene oxide and did not metabolize limonene. Pseudomonas fluorescens was a more versatile micro‐organism and metabolized limonene in two different ways. The first (anaerobic, cofactor‐independent, noninducible) allowed limonene elimination by synthesizing α‐terpineol. The second (aerobic, cofactor‐dependent) involved limonene‐1,2‐oxide as an intermediate for energy production through a β‐oxidation process. Significance and Impact of the Study: Enzymatic isomerization of β‐ to α‐pinene was described for the first time for both strains. Alpha‐terpineol production by P. fluorescens was very efficient and appeared as a promising alternative for the commercial production of this bioflavour.     

Biomass production of Cymbopogon citratus (D.C.) stapf., a medicinal plant, in temporary immersion systems

Summary In vitro plants of lemon grass were established, starting from shoot apices derived from plants cultivated under field conditions. The effect of the immersion frequency (two, four, and six immersions per day) on the production of biomass in temporary immersion systems (TIS) of 1 liter capacity was studied. The highest multiplication coefficient (12.3) was obtained when six immersions per day were used. The maximum values of fresh weight (FW; 62.2 and 66.2 g) were obtained with a frequency of four and six immersions per day, respectively. However, the values for dry weight (DW; 6.4g) and height (8.97cm) were greater in the treatment with four immersions per day. The TIS used in this work for the production of lemon grass biomass may offer the possibility of manipulating the culture parameters, which can influence the production of biomass and the accumulation of secondary metabolites. We describe for the first time the in vitro production of Cymbopogon citratus biomass in TIS.     

The potency of lemon (Citrus limon L.) essential oil to control some fungal diseases of grapevine wood

This study aimed to evaluate the in vitro antifungal activity (AA) of the essential oil (EO) of lemon (Citrus limon L.) against three pathogenic fungi attacking grapevine wood. The composition of the EO was also studied. Ten volatile components were identified by gas chromatography–mass spectrometry. The results showed that the EO consists of volatile components where monoterpene hydrocarbons are the most abundant ones. Four major components were identified, which represent 99.9% of the total EO (limonene, neral, ß-pinene, and ?-terpinene). The AA of the EO was evaluated against three pathogenic fungi attacking grapevine wood (Eutypa sp., Botryosphaeria dothidea, and Fomitiporia mediterranea). The results showed that the EO exerts AA against all tested fungi and significantly inhibits their growth. Eutypa sp. is the most sensitive fungus. These results show, for the first time, a new use for the EO of lemon (C. limon L.) to control fungal diseases of grapevine wood.     

Composition, antimicrobial and antioxidant activity of the extracts of Eryngium palmatum Pančić and Vis. (Apiaceae)

The chemical composition, antimicrobial and antioxidant activity of Eryngium palmatum, an endemic plant species from the Balkan Peninsula, were investigated. The flavonoids apigenin (9.5±0.3 mg g^?1) and apigenin 7-O-glucoside (2.4±0.1 mg g^?1) were determined in a methanol extract of aerial parts using HPLC analysis. The methanol extract of roots contained catechin (5.0±0.1 mg g^?1), epicatechin (2.9±0.1 mg g^?1), chlorogenic acid (1.6±0.0 mg g^?1), gallic acid (0.9±0.0 mg g^?1) and rosmarinic acid (0.9±0.2 mg g^?1). GC-FID and GCMS analysis of a chloroform extract of aerial parts showed that the main volatile constituents were falcarinol, linoleic acid, hexadecanoic acid and methyl linoleate (comprising 32.6%; 24.4%; 19.9; 13.2% of the volatile fraction, respectively), while octanoic acid, tetradecanol and dodecanol dominated in the chloroform extract of the roots (34.9%; 25.8%; 22.2% of the volatile fraction, respectively). Investigation of antimicrobial activity by broth microdilution showed that the methanol and chloroform extracts of aerial parts and roots exerted a significant effect (MIC 3.5–15.6 μg mL^?1) against tested Gram-positive and Gram-negative bacteria. The methanol extracts of aerial parts or roots exerted moderate ferric reducing antioxidant power, DPPH radical scavenging activity and hydroxyl radical scavenging activity.     

Phenolic composition and antioxidant properties of some traditionally used medicinal plants affected by the extraction time and hydrolysis

Introduction – Polyphenolic phytochemicals in traditionally used medicinal plants act as powerful antioxidants, which aroused an increasing interest in their application in functional food development. Objective – The effect of extraction time (5 and 15 min) and hydrolysis on the qualitative and quantitative content of phenolic compounds and antioxidant capacity of six traditionally used medicinal plants (Melissa officinalis L., Thymus serpyllum L., Lavandula officinalis Miller, Rubus fruticosus L., Urtica dioica L., and Olea europea L.) were investigated. Methodology – The content of total phenols, flavonoids, flavan‐3‐ols and tannins was determined using UV/Vis spectrophotometric methods, while individual phenolic acids, flavones and flavonols were separated and detected using HPLC analysis. Also, to obtain relevant data on the antioxidant capacity, two different assays, (2,2‐azino‐bis(3‐ethylbenzthiazoline‐6‐sulphonic acid) (ABTS) radical scavenging and ferric reducing/antioxidant power (FRAP) assays were used. Results – The extraction efficiency of phenolics, as well as the antioxidant capacity of plant extracts, was affected by both prolonged extraction and hydrolysis. The overall highest content of phenolic compounds was determined in hydrolyzed extract of blackberry leaves (2160 mg GAE/L), followed by the non‐hydrolyzed extract of lemon balm obtained after 15 min of extraction (929.33 mg GAE/L). The above extracts also exhibited the highest antioxidant capacity, while extracts of olive leaves were characterized with the lowest content of phenolic compounds, as well as the lowest antioxidant capacity. The highest content of rosmarinic acid, as the most abundant phenolic compound, was determined in non‐hydrolyzed extract of lemon balm, obtained after 15 min of extraction. Although the hydrolysis provided the highest content of polyphenolic compounds, longer extraction time (15 min) was more efficient to extract these bioactives than shorter extraction duration (5 min). Conclusion – The distribution of detected phenolic compounds showed a wide variability with regard to their botanical origin. Examined medicinal plants showed to be a valuable supplement to a daily intake of bioactive compounds. Copyright © 2010 John Wiley & Sons, Ltd.     

Toxicity of citrus essential oils against Ceratitis capitata (Diptera: Tephritidae) larvae

Citrus peel essential oils are considered to constitute the most important resistance factor of citrus fruits against fruit flies. Essential oils were obtained from three sweet orange varieties, one bitter orange and one lemon variety. Yield, chemical composition and toxicity against neonates of the Mediterranean fruit fly were determined. Based on chemical analysis, the toxicity of commercially purchased major and minor components (monoterpenes and sesquiterpenes) of essential oils was determined. In addition, fractions were prepared to evaluate the role of minor components in the toxicity of crude essential oils. Limonene was by far the most abundant ingredient (96.2–97.4%) in all sweet orange varieties and in bitter orange, while the concentration of limonene was much lower in lemon essential oils (74.3%). Orange and bitter orange essential oils were more toxic than lemon essential oils. The toxicity of orange and bitter orange essential oils was similar to that of their major component limonene. In tests of commercially purchased chemicals, the oxygenated components of essential oils were more toxic than hydrocarbons but their low concentration in citrus essential oils could not affect the toxic activity of essential oils. The presence of α-pinene and β-pinene seems to account for the lower toxicity of lemon essential oils in relation to other citrus essential oils. The importance of understanding the toxicity of essential oils in relation to their composition and their role regarding the resistance of citrus fruits to Ceratitis capitata infestation is discussed.     

Solid-phase microextraction for determining twelve orange flavour compounds in a model beverage emulsion

Solid-phase microextraction (SPME) coupled to gas chromatography has been applied for the headspace analysis (HS) of 12 target flavour compounds in a model orange beverage emulsion. The main volatile flavour compounds studied were: acetaldehyde, ethyl acetate, alpha-pinene, ethyl butyrate, beta-pinene, myrcene, limonene, gamma-terpinene, octanal, decanal, linalool and citral (neral plus geranial). After screening the fibre type, the effect of other HS-SPME variables such as adsorption temperature (25-55 degrees C), extraction time (10-40 min), sample concentration (1-100% w/w), sample amount (5-10 g) and salt amount (0-30% w/w) were determined using a two-level fractional factorial design (2(5-2)) that was expanded further to a central composite design. It was found that an extraction process using a carboxen-polydimethylsiloxane fibre coating at 15 masculineC for 50 min with 5 g of diluted emulsion 1% (w/w) and 30% (w/w) of sodium chloride under stirring mode resulted in the highest HS extraction efficiency. For all volatile flavour compounds, the linearity values were accurate in the concentration ranges studied (r(2) > 0.97). Average recoveries that ranged from 90.3 to 124.8% showed a good accuracy for the optimised method. The relative standard deviation for six replicates of all volatile flavour compounds was found to be less than 15%. For all volatile flavour compounds, the limit of detection ranged from 0.20 to 1.69 mg/L.     

Bioremediation of Aroclor 1242 by a consortium culture in marine sediment microcosm

Plant terpenes have proven to be effective in stimulation of polychlorinated biphenyls (PCBs) biodegradation in soil systems. However, data on the application of plant terpenes in marine sediments contaminated with PCBs remains limited. The aim of this study was to ascertain the roles of a PCB degrading consortium and plant terpenes in stimulation of PCB biodegradation in marine sediments. The consortium culture 1-2Mix (strains 1-2M and 1-2T in commensalism), a utilizer of biphenyl and a natural substrate was enriched and isolated from marine sediments from the Busan coast, South Korea. PCB degradation by this culture was shown to be more effectively induced by tangerine peel extract than other known substrates (limonene, pinene, and cymene). Coastal sediment microcosms inoculated with 1-2Mix were set up to elucidate the effect of the consortium and plant terpenes on degradation of Aroclor 1242. After four weeks, the highest removal rates of PCBs, compared with the control (autoclaved sediment and no inoculation of 1-2Mix), were observed in order of the inducers tested; biphenyl (71.1%), tangerine peel extract (69.5%), surfactant (66.0%), and limonene (63.0%). Bioaugmentation effect was doubled in the presence of natural substrates such as tangerine peel extract and limonene, indicating effectiveness of these substrates in biostimulation. It was concluded that the tangerine peel extract could replace biphenyl as a feasible induction substrate for effective remediation of PCBs in the marine sediment.     

Single cell protein production from mandarin orange peel

Summary As the hydrolysis of mandarin orange peel with macerating enzyme (40°C, 24 h) produced 0.59 g g^–1 reducing sugar per dry peel compared to 0.36 by acid-hydrolysis (15 min at 120°C with 0.8 N H₂SO₄), the production of single cell protein (SCP) from orange peel was studied mostly using enzymatically hydrolyzed orange peel.When the enzymatically hydrolyzed peel media were used, the utilization efficiency of reducing sugars (%) and the growth yield from reducing sugars (g g^–1) were: 63 and 0.51 for Saccharomyces cerevisiae; 56 and 0.48 for Candida utilis; 74 and 0.69 for Debaryomyces hansenii and 64 and 0.70 for Rhodotorula glutinis. SCP production from orange peel by D. hansenii and R. glutinis were further studied. Batch cultures for 24 h at 30°C using 100 g dried orange peel produced 45 g of dried cultivated peel (protein content, 33%) with D. hansenii and 34 g (protein content, 50%) with R. glutinis, and 38 g (protein content, 44%) with a mixture of both yeasts.     

Terpene down-regulation in orange reveals the role of fruit aromas in mediating interactions with insect herbivores and pathogens

Plants use volatile terpene compounds as odor cues for communicating with the environment. Fleshy fruits are particularly rich in volatiles that deter herbivores and attract seed dispersal agents. We have investigated how terpenes in citrus fruit peels affect the interaction between the plant, insects, and microorganisms. Because limonene represents up to 97% of the total volatiles in orange (Citrus sinensis) fruit peel, we chose to down-regulate the expression of a limonene synthase gene in orange plants by introducing an antisense construct of this gene. Transgenic fruits showed reduced accumulation of limonene in the peel. When these fruits were challenged with either the fungus Penicillium digitatum or with the bacterium Xanthomonas citri subsp. citri, they showed marked resistance against these pathogens that were unable to infect the peel tissues. Moreover, males of the citrus pest medfly (Ceratitis capitata) were less attracted to low limonene-expressing fruits than to control fruits. These results indicate that limonene accumulation in the peel of citrus fruit appears to be involved in the successful trophic interaction between fruits, insects, and microorganisms. Terpene down-regulation might be a strategy to generate broad-spectrum resistance against pests and pathogens in fleshy fruits from economically important crops. In addition, terpene engineering may be important for studying the basic ecological interactions between fruits, herbivores, and pathogens.     

Oviposition behaviour and larval development of Anastrepha fraterculus from Argentina in citrus

Citrus peel physicochemical attributes are considered the main components conferring partial or even total resistance to fruit fly (Diptera: Tephritidae) infestation. Fruit fly females adapt their ovipositional strategies to overcome such resistance. Here, we explored the effects of citrus species (Rutaceae) on the ovipositional behaviour of the South American fruit fly, Anastrepha fraterculus (Wiedemann), and on its immature development. Particularly, we investigated the effects of (1) citrus species on oviposition behaviour and immature development, (2) citrus species on oviposition preference and on the location of the eggs at different depth in the citrus peel, and (3) harvest season and post‐harvest storage time on oviposition behaviour and immature development in lemon. Citrus species influenced ovipositional behaviour and affected survival of immature stages. Females laid eggs in lemon [Citrus limon (L.) Burm.], orange [Citrus sinensis (L.) Osbeck], and grapefruit (Citrus paradisi Macfadyen). In orange and lemon, larvae were found dead close to the oviposition areas, suggesting chemically mediated resistance mechanisms. Under choice conditions, females preferred grapefruit over lemon and bigger clutches were found in the layers where embryonic development is favoured. Unsuitability of lemon as a medium to complete development was neither affected by harvest season nor by storage time of the fruit after harvest. The physical and chemical characteristics of the peel were distinctive to each citrus species and may have affected the specific levels of resistance of these citrus species to infestation by A. fraterculus.     

Preparation of cation exchanger from lemon and sorption of divalent heavy metals

A cation exchanging material was developed from lemon by modifying the pectic-cellulosic substances in the lemon peel by lemon juice having citric acid. For this purpose, chopped lemon removed from seeds and yellow skin was heated in two stages, firstly at 50 degrees C for 24h and subsequently at 120 degrees C for 2h. The material obtained was ground, repeatedly washed with water and dried. Lemon peel and lemon resin obtained were characterized through physicochemical analyses and FTIR spectroscopy. Heavy metal binding performance of this material was determined by removal tests conducted by using 10mM solutions of divalent metals. Experimental results show that the resin prepared from lemon is effective especially for Pb and Cu removals. For a lemon resin dosage of 10 g l(-1), sorption affinity of divalent metal ions is found to be in an order of Pb>Cu>Ni>Fe>Cd>Zn>Co>Mn. Typically, sorption capacities are about 0.87 and 0.43 mmol g(-1) for Pb and Mn, respectively.     

The effect of imazalil in the control of decay in yellow yam caused by Penicillium sclerotigenum

A strain of Penicillium sclerotigenum isolated from decaying yellow yams (Dioscorea cayenensis) was found to have developed resistance to benzimidazole fungicides and the use of imazalil was investigated as an alternative agent for controlling it. Two formulations were tested and proved to be equally effective in controlling decay at a concentration of 50 mg imazalil litre^-1 if the yams were dipped in it for 5 s; concentrations down to 10 mg litre^-1 were effective if the immersion time was increased to 5 min or more. These treatments gave good control of decay when applied up to 24 h after inoculation but were less effective when application was 48 h after inoculation, although at 500 mg litre^-1 there was some indication that levels of decay were decreased when compared with untreated tubers. Fungal penetration was unaffected or increased by increasing delays in the time of the fungicide application depending on the concentration applied. In trial shipments of yams from Jamaica to Britain imazalil alone was less effective in controlling overall decay because of the presence of a Rhizopus species. This could be controlled by the addition of dichloran to the dip. Imazalil residues were highest in the peel; levels in the peeled tuber varied and tended to decrease during storage but were generally in the range of 0·5 - 0·8 mg kg^-1 which is higher than the maximum levels accepted for other produce. A simple cooking test showed that residues in treated yam slices declined by about one third after boiling the slices in water for 45 min.     

Exploring non-host plant-based management strategy with lemongrass,garlic and guava volatiles for the African citrus triozid

The African citrus triozid (ACT), Trioza erytreae, is an important pest of citrus. Both nymphs and adults damage the plant by feeding on the sap causing young shoots to die. Trioza erytreae also vectors Candidatus Liberibacter africanus, the bacteria that cause citrus greening disease. Since certain non-host plants are known to repel insect pests, it is important to investigate how such plants can be exploited to manage T. erytreae. Here, we screened effects of odours of three non-host plants namely guava (Psidium guajava), garlic (Allium sativum) and lemongrass (Cymbopogon citratus) against T. erytreae's location of a common host plant, rough lemon (Citrus jambhiri) and showed that repellence varied interspecifically with the plants. Using cage assays, we found that guava and garlic decreased the attraction of females but not males of T. erytreae to rough lemon volatiles. Chemical analysis by coupled gas chromatography/mass spectrometry (GC/MS) showed that volatiles of three of the plants were dominated by terpenoids; guava (69% comprised of limonene, 34%, (E)-β-ocimene, 29% and (Z)-β-Ocimene, 6%), lemongrass (56% comprised of geranial, 26%, neral, 19% and myrcene, 11%) and rough lemon (74% comprised of limonene, 53%, sabinene, 11% and (E)-β-ocimene, 10%). On the other hand, the volatile profile of garlic was dominated by benzenoids and saturated compounds (85% comprised of benzaldehyde, 12%, benzyl alcohol, 17%, nonanal, 31%, decanal, 13% and hexadecane, 12%). Our results suggest that non-host plant volatile composition and richness in specific compounds may contribute to influencing T. erytreae response to its host, with garlic and guava as potential non-host plants that can be exploited in the management of the pest.     

Seasonal emission of monoterpenes by the Mediterranean tree Quercus ilex in field conditions: Relations with photosynthetic rates, temperature and volatility

The relationships of monoterpene emission with temperature, light, photosynthesis and stomatal conductance (g_s) were studied in Quercus ilex L. trees throughout the four annual seasons under field conditions. The highest monoterpene emission was measured in spring and summer (midday average of 11 μg [g DW]^?1 h^?1), whereas the lowest rates were found in autumn and winter (midday averages of 0.51 and 0.23 μg [g DW]^?1 h^?1, respectively). In spring and summer, limonene was the monoterpene emitted at highest rate (midday averages of 5.27–6.69 μg [g DW]^?1 h^?1), whereas α-pinene was emitted the most in autumn and winter (midday averages of 0.31 μg [g DW]^?1 h^?1). The monoterpenes limonene, α-pinene and β-pinene represented about 75–95% of total detected monoterpenes. The total monoterpene emission rates represented about 0.04% of carbon fixed in autumn, 0.17% in winter, 0.84–2.51% in spring and 1.22–5.13% in summer. Significant correlations of total monoterpene emission with temperature were found when considering either summer emission or the emission over the entire year, whereas significant correlations with net photosynthetic rates were only found when considering summer season. Among individual terpenes, the most volatile, α-pinene and β-pinene, were more correlated with temperature than with net photosynthetic rates whereas the less volatile limonene was more correlated with net photosynthetic rate. Thus, under field conditions it seems that dependency of monoterpene emission on photosynthetic rate or temperature is partly related with volatility of the compounds. Influences of seasonality, temperature, photosynthetic rates and volatility should be considered in inventories and models of emission rates in Mediterranean ecosystems.     

A lemon myrtle extract inhibits glucosyltransferases activity of Streptococcus mutans

Streptococcus mutans is a bacterium found in human oral biofilms (dental plaques) that is associated with the development of dental caries. Glucosyltransferases (GTFs) are key enzymes involved in dental plaque formation, and compounds that inhibit their activities may prevent dental caries. We developed a screening system for GTF-inhibitory activities, and used it to profile 44 types of herbal tea extracts. Lemon myrtle (Backhousia citriodora) extract exhibited the highest GTF-inhibitory activity, with an IC₅₀ for GTF in solution of 0.14 mg mL^?1. Furthermore, lemon myrtle extracts had the third-highest polyphenol content of all tested extracts, and strongly inhibited S. mutans biofilm. Interestingly, lemon myrtle extracts did not inhibit cell growth.     

Quantification of protopectinase SE, an endopolygalacturonase with pectin-releasing activity from Geotrichum klebahnii

Pectin releasing activity of protopectinase SE (PPase-SE) from Geotrichum klebahnii (= G. penicillatum = Trichosporon penicillatum) ATCC 42397 was determined using different batches of lemon protopectin as substrate. Results obtained showed a high degree of variability depending on the batch of protopectin used. As PPase-SE also shows polygalacturonase (PGase) activity, a method for the assay of this activity was optimised. The best assay conditions were: substrate (polygalacturonic acid) concentration of 2.0 g 1^–1, reaction time of 10 min and up to 0.17 PGase units per test tube.