Antioxidant, antibacterial, and antiviral effects of Lactuca sativa extracts

Antioxidant, antibacterial and antiviral effects of aqueous and methanol extracts of Lactuca sativa var longifolia leaves were investigated. The antioxidant activity was evaluated using the DPPH assay. The effect of the extracts against 5 Gram-positive and 6 Gram-negative bacteria was tested. The antiviral activity was determined against human cytomegalovirus (HCMV) strain AD-169 (ATCC Ref. VR 538) and coxsackie B virus type 3 (CoxB-3) using a cytopathic effect (CPE) reduction assay. The methanol extract had the highest total phenolic contents (235.31 mg CE/g extract). It exhibited a significantly (p < 0.05) greater hydroxyl radical-scavenging activity (IC₅₀ = 3.5 μg/ml) than the aqueous extract (4.1 μg/ml). It was also the most effective extract with the lowest MIC (2.5 mg/ml) against all Gram negative and Gram positive bacteria. Methanol and aqueous extracts exhibited antiviral activity against HCMV and Cox-B3 viruses with IC₅₀ of 200 μg/ml.     

Baseline and differential sensitivity to mandipropamid among isolates of Peronophythora litchii, the causal agent of downy blight on litchi

Litchi downy blight caused by Peronophythora litchii is a devastating disease of litchi plants in China. Control of litchi downy blight requires numerous fungicide applications. A new carboxylic acid amide (CAA) fungicide, mandipropamid, was examined for its in vitro effects on multiple asexual stages of four single-sporangium P. litchii isolates and protective activity against downy blight in detached fruit assays. Though mandipropamid did not affect discharge of zoospores from sporangia, it strongly inhibited mycelial growth (mean EC₅₀ = 0.0048 μg ml⁻¹), sporangia production (mean EC₅₀ = 0.0032 μg ml⁻¹), germination of encysted zoospores (mean EC₅₀ = 0.0023 μg ml⁻¹), and germination of sporangia (mean EC₅₀ = 0.0061 μg ml⁻¹). On detached fruit, 0.39, 1.56 and 6.25 μg ml⁻¹ of mandipropamid were superior in reducing downy blight compared to metalaxyl and flumorph, however, the 25 μg ml⁻¹ application rate was necessary for all three CAA fungicides to completely inhibit the disease. In 2007, 100 isolates from Fujian, Guangdong, and Guangxi Provinces of China were characterized for the baseline sensitivity to mandipropamid. The isolates obtained from different provinces showed similar baseline sensitivities to mandipropamid. Baseline sensitivities formed a unimodal curve with mean EC₅₀ values of 0.0055 ± 0.0012 μg ml⁻¹ for inhibition of mycelial growth. The described baseline sensitivities of P. litchii populations will be useful for monitoring possible shifts in sensitivity to mandipropamid.     

Effects of crude seed extracts of Annona atemoya and Annona squamosa L. against the cabbage looper, Trichoplusia ni in the laboratory and greenhouse

Antifeedant, growth inhibitory and toxic effects of crude seed extracts of Annona squamosa and Annona atemoya from Fazenda Viveiro Bona, Parasisópolis – Minas Gerais, Brazil, were evaluated against the cabbage looper, Trichoplusia ni (Hübner) (Lepidoptera: Noctuidae) using different bioassays. Crude methanolic seed extracts deterred feeding of third instar T. ni larvae in a leaf disc choice bioassay. A. squamosa was ∼10 times more active as a feeding deterrent than A. atemoya (DC₅₀ = 2.3 mg/ml vs. 20.1 mg/ml). A. squamosa was ∼three times more active as a growth inhibitor than A. atemoya (EC₅₀ = 38.0 ppm vs. 117.0 ppm). Methanolic seed extracts of A. squamosa and A. atemoya were toxic to third instar T. ni larvae both through topical and oral application. A. squamosa was more toxic through feeding (LC₅₀ = 167.5 ppm vs. 382.4 ppm) whereas, A. atemoya exerted greater toxicity via topical application (LC₅₀ = 301.3 μg/larva vs. 197.7 μg/larva). Both A. squamosa and A. atemoya extracts reduced leaf area consumption and larval growth in a greenhouse experiment. Our results indicate that both A. squamosa and A. atemoya have potential for development as botanical insecticides, especially for local use in Brazil.     

Sensitivity of Botryosphaeria dothidea from apple to tebuconazole in China

The white rot disease caused by Botryosphaeria dothidea threatens apple production in Bohai bay area and along the Yellow River of China, where disease control is largely dependent on fungicides such as tebuconazole. A total of 146 isolates of B. dothidea obtained from different apple orchards in six provinces were tested for their sensitivity to tebuconazole, carbendazim and iprodione. The EC₅₀ values of all tested isolates for tebuconazole were from 0.035 to 1.415 μg/mL. The broad range of EC₅₀ values of tebuconazole suggests an obvious variation among the 146 isolates. Isolate HB13 (EC₅₀ = 1.415 μg/mL) showed reduced sensitivity to tebuconazole, with an EC₅₀ value significantly higher than those of the other 145 isolates tested. The low sensitivity of HB13 was stable after 15 generations, and this isolate showed similar pathogenicity as susceptible strains. EC₅₀ correlation analysis indicates no cross resistance between tebuconazole and carbendazim and iprodione. Field efficacy trials showed that tebuconazole remains very effective for apple white rot control in China.     

Toxicity of quinones against two-spotted spider mite and three species of aphids in laboratory and greenhouse conditions

Toxicities of the eight quinones were evaluated through leaf dip bioassays conducted against Tetranychus urticae, Myzus persicae, Myzocallis walshii, and Illinoia liriodendri. Based on LC₅₀ values, plumbagin (LC₅₀ = 0.001%) was the most active compound against T. urticae and ubiquinone Q₀ (LC₅₀ = 0.005%), plumbagin (LC₅₀ = 0.010%), and dibromothymoquinone (LC₅₀ = 0.012%) were the most active compounds against M. persicae. The most active compounds against M. walshii were juglone (LC₅₀ = 0.011%) and ubiquinone Q₀ (LC₅₀ = 0.019%), whereas dibromothymoquinone (LC₅₀ = 0.030%), plumbagin (LC₅₀ = 0.033%) and ubiquinone Q₀ (LC₅₀ = 0.058%) were the most toxic to I. liriodendri. Ecotrol (positive control) was the least toxic compound (LC₅₀ = 0.39%) against T. urticae and M. persicae (LC₅₀ = 0.447%). Although the majority of the compounds tested were toxic to all four test species in residual bioassays, there was little overlap among the test species in terms of susceptibility to the compounds and interspecific differences were observed. Regarding structure-activity relationships for quinones, the addition of a hydroxyl group resulted in a significant increase in the toxicity of the 1,4-naphthoquinones, and those possessing a methyl group exhibited the highest levels of activity in T. urticae. The bromine atom at the 2- and 5-positions of the benzoquinone ring is crucial to the toxicity of the compounds against I. liriodendri. Toxicity was greatly affected not only by the number of hydroxyl groups, but also by their positions in the ring in the case of M. walshii. Juglone and plumbagin as residual toxins in the laboratory also reduced the population of two-spotted spider mites compared to EcoTrol™ (positive control) and the negative control in the greenhouse experiment. Some quinones tested may have potential as commercial insecticides and miticides, or alternatively, could serve as lead compounds for the development of more potent crop protection agents.     

Maritime Halophyte Species from Southern Portugal as Sources of Bioactive Molecules

Extracts of five halophytes from southern Portugal (Arthrocnemum macrostachyum, Mesembryanthemum edule, Juncus acutus, Plantago coronopus and Halimione portulacoides), were studied for antioxidant, anti-inflammatory and in vitro antitumor properties. The most active extracts towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were the methanol extracts of M. edule (IC₅₀ = 0.1 mg/mL) and J. acutus (IC₅₀ = 0.4 mg/mL), and the ether extracts of J. acutus (IC₅₀ = 0.2 mg/mL) and A. macrostachyum (IC₅₀ = 0.3 mg/mL). The highest radical scavenging activity (RSA) against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical was obtained in the ether extract of J. acutus (IC₅₀ = 0.4 mg/mL) and H. portulacoides (IC₅₀ = 0.9 mg/mL). The maximum total phenolic content (TPC) was found in the methanol extract of M. edule (147 mg gallic acid equivalents (GAE)/g) and in the ether extract of J. acutus (94 mg GAE/g). Significant decreases in nitric oxide (NO) production were observed after incubation of macrophages with lipopolysaccharide (LPS) and the chloroform extract of H. portulacoides (IC₅₀ = 109 µg/mL) and the hexane extract of P. coronopus (IC₅₀ = 98.0 µg/mL). High in vitro cytotoxic activity and selectivity was obtained with the ether extract of J. acutus. Juncunol was identified as the active compound and for the first time was shown to display selective in vitro cytotoxicity towards various human cancer cells.     

Sensitivity of Ascochyta rabiei populations to prothioconazole and thiabendazole

Ascochyta rabiei causes Ascochyta blight, a yield-limiting disease of chickpea (Cicer arietinum) world-wide. In 2007, fungal populations of A. rabiei resistant to the QoI group of fungicides were detected in the Northern Great Plains of the United States. Assays were conducted to determine fungal sensitivity for two alternative fungicidal modes of action. A total of 78 isolates of A. rabiei collected between 1983 and 2007 were screened to determine baseline sensitivity to the demethylation-inhibiting foliar fungicide, prothioconazole, and 100 isolates collected between 1987 and 2007 were screened for sensitivity to the methyl benzimidazole carbamate (MBC) fungicide, thiabendazole. Isolates were tested using an in vitro mycelial growth assay to determine the effective fungicide concentration at which 50% of fungal growth was inhibited (EC₅₀) for each isolate-fungicide combination. Baseline EC₅₀ values of prothioconazole ranged from 0.0526 to 0.2958 μg/ml, with a mean of 0.1783 μg/ml. Isolates of A. rabiei collected from 2007 to 2009 from North Dakota chickpea fields exposed to prothioconazole, were screened for prothioconazole sensitivity using the same assay. Mean EC₅₀ values for these isolates were 0.3544 μg/ml, 0.3746 μg/ml, and 0.7820 μg/ml, respectively. These values represent an approximate 2.0 (2007-2008) and 4.4-fold (2009) decrease in sensitivity from the baseline mean. EC₅₀ values of thiabendazole ranged from 1.192 to 3.819 μg/ml, with a mean of 2.459 μg/ml. No significant decrease in fungicide sensitivity was observed for thiabendazole. To date, no loss of Ascochyta blight control has been observed with the use of either prothioconazole or thiabendazole.     

Antifungal activity of polyacetylenes isolated from Cirsium japonicum roots against various phytopathogenic fungi

Antifungal substances from a methanol extract of Cirsium japonicum roots were purified and characterized, and their antifungal activities against various plant pathogens were evaluated. Three polyacetylene substances were isolated from roots of C. japonicum using repeated column chromatography; these were identified as ciryneol A, ciryneol C and 1-heptadecene-11,13-diyne-8,9,10-triol by mass and nuclear magnetic resonance spectral analyses. In vitro antifungal activity of the three substances varied according to compound and target species. Magnaporthe oryzae, Colletotrichum coccodes, Colletotrichum acutatum, Pythium ultimum and Botrytis cinerea were relatively sensitive to the three polyacetylenes, with IC₅₀ values below 50 μg mL⁻¹. In vivo, they all showed similar and broad antifungal spectra against the seven plant diseases tested. At 500 μg mL⁻¹, all three compounds effectively suppressed the development of rice blast, rice sheath blight, tomato late blight, wheat leaf rust and red pepper anthracnose, with control values over 90%. They were highly active especially against wheat leaf rust; they controlled the development of this disease more than 88% even at a concentration of 125 μg mL⁻¹. In addition, ciryneol C effectively suppressed barley powdery mildew. This is the first report on the antifungal activities of the three polyacetylenes from roots of C. japonicum against plant pathogenic fungi. Polyacetylenes from roots of C. japonicum may contribute to the development of environmentally safer alternatives to protect crops from various phytopathogenic fungi.     

Bioactive properties and chemical constituents of methanolic extract and its fractions from Jatropha curcas oil

The present work is designed to evaluate the bioactive properties of the crude methanolic extract of Jatropha curcas oil and its solvent fractions. The crude methanolic extract obtained was fractionated using a hydrophilic lipophilic balanced (HLB) cartridge and then eluted with different solvents in the order of hexane (F1), dichloromethane (F2), chloroform (F3), ethyl acetate (F4) and methanol (F5), respectively. Total phenolic content of the crude methanolic extract and its fractions was in the range of 0.19-4.5 mg/g as gallic acid equivalent. Antioxidant activity of the crude methanolic extract and its fractions were determined by two complementary test methods, namely, phenanthroline method and 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging method. All samples demonstrated weak antioxidant activity (150-851 μmol Fe/100 g of the extract and IC₅₀ of 1.05-13.5 mg/mL). When compared to butylated hydroxytoluene (BHT), a reference synthetic antioxidant, both showed weaker antioxidative potential. The evaluation of antimicrobial activity of the extracts was performed using a disc diffusion method and a micro-well dilution method against six economic plant disease bacteria. The results showed that all extracts possessed strong to moderate antibacterial activity with varying degrees of growth inhibition against the test bacteria. The minimum inhibitory concentrations (MIC) were in the range of 14.92-428.6 μg/mL. In addition, the chemical constituents in each fraction of the extract were subjected to analyze by gas chromatography-mass spectrometry (GC-MS). The eleven constituents were identified. Among them, 2,4-di-tert-butylphenol, methyl 3-(3,5-di-tert-butyl-4-hydroxyphenyl)propionate and linoleic acid may be the main cause of its strong antibacterial activity. Therefore, this oil present in the methanolic extract had great potential as effective antibacterial sources.     

Arbutus unedo L. leaves as source of phytochemicals with bioactive properties

In recent years the strawberry tree (Arbutus unedo L.) is being gradually replaced by other species with higher economic value. With the ultimate goal of selecting superior genotypes, the present work was initiated to study the antioxidant and antimicrobial activities, and total phenolic content in 19 different genotypes of A. unedo leaves from the Trás-os-Montes region of Portugal.The genotype Bragança 1 contains higher total phenolic content (215.0 mg GAE/g_extract) whereas the Vila Boa 4 genotype shows lower total phenolic content (148.0 mg GAE/g_extract). In both methods tested to evaluate the antioxidant activity, Vila Verde and Donai displayed the highest antioxidant capacity (EC₅₀ values of 0.088 and 0.090 mg/mL, respectively, for DPPH; EC₅₀ values of 0.233 and 0.245 mg/mL, respectively, for reducing power assay) while Vila Boa 2 reported the lowest antioxidant potential (EC₅₀ values of 0.142 and 0.378 mg/mL, respectively, in DPPH and reducing power methods). Linear negative correlations were established between the total phenol contents and the EC₅₀ values for both of the antioxidant activity methods tested. Preliminary assays for antimicrobial potential showed that extracts from A. unedo leaves display antibacterial activity, with MIC values of 1 and 5 mg/mL for some Gram-positive and Gram-negative bacteria, respectively. Taken together, the results suggest that A. unedo leaves are a potential source of natural compounds with valuable bioactive properties that could be explored by the pharmaceutical, chemical and food industries.     

Comparative assessment of antioxidant and cholinesterase inhibitory properties of the marigold extracts from Calendula arvensis L. and Calendula officinalis L.

Calendula sp. is an important medicinal and industrial plant with various bioactivities. In this study, we examined enzyme inhibitory effects of the n-hexane, dichloromethane, acetone, ethyl acetate, methanol, and water extracts of the leaf and flowers of Calendula arvensis L. and C. officinalis L. against acetylcholinesterase (AChE) and butyrylcholinesterase (BChE). The extracts were screened for their antioxidant activity using 2,2-diphenyl-1-picrylhydrazyl (DPPH), ferric ion-chelating capacity and ferric-reducing antioxidant power (FRAP) assays at 250, 500, and 1000 μg mL⁻¹. Total phenol and flavonoid quantification of the extracts was achieved using Folin-Ciocalteau and AlCl₃ reagents, respectively. The ethyl acetate extract of C. arvensis flowers was the most active in AChE inhibition assay (31.24 ± 1.29%), while the n-hexane extract of C. officinalis leaves exerted the highest ferric ion-chelating capacity (74.27 ± 2.25%). Thin layer chromatographic analysis indicated presence of flavonoid and triterpene derivatives mainly in the extracts.     

Antifungal and antiaflatoxigenic efficacy of Caesulia axillaris Roxb. essential oil against fungi deteriorating some herbal raw materials, and its antioxidant activity

The study deals with evaluation of antifungal and antiaflatoxigenic Caesulia axillaris Roxb. essential oil (EO) against herbal raw materials deteriorating fungi and its free radical scavenging activity. During mycoflora analysis these herbal raw materials were found to be severely contaminated by different fungi and aflatoxins. A total of nine different fungal species were isolated from three herbal raw materials. Aspergillus flavus LHPtc was recorded as the highest aflatoxin B1 producing strain. EOs of some plants were tested for their fungitoxicity against the toxigenic strain A. flavus LHPtc, and C. axillaris EO was found as potent fungitoxicant. C. axillaris EO was chemically characterized through GC-MS analysis which depicted the presence of 18 compounds, dl-limonene and Euasarone being the major components. The EO exhibited broad spectrum of fungitoxicity against fungi causing postharvest deterioration of herbal raw materials. At 1.0 μl ml⁻¹ the oil showed complete inhibition of fungal growth and aflatoxin B₁ production was inhibited at 0.8 μl ml⁻¹. Free radical scavenging activity of the oil was also recorded by 2,2-diphenyl-1-picrylhydrazyl assay, and its IC₅₀ value was found 18 μl ml⁻¹. The safety limit of the EO was determined in terms of LD₅₀ on mice, which was 9166.6 μl kg⁻¹, suggesting its non mammalian toxicity. The EO of C. axillaris may be recommended as a plant based preservative in enhancement of shelf life of herbal raw materials by preventing their lipid peroxidation as well as biodeterioration due to fungal and aflatoxin contamination.     

Metabolic Profiling and In Vitro Assessment of the Biological Activities of the Ethyl Acetate Extract of Penicillium chrysogenum “Endozoic of Cliona sp. Marine Sponge” from the Red Sea (Egypt)

Marine sponge-derived endozoic fungi have been gaining increasing importance as promising sources of numerous and unique bioactive compounds. This study investigates the phytochemical profile and biological activities of the ethyl acetate extract of Penicillium chrysogenum derived from Cliona sp. sponge. Thirty-six compounds were tentatively identified from P. chrysogenum ethyl acetate extract along with the kojic acid (KA) isolation. The UPLC-ESI-MS/MS positive ionization mode was used to analyze and identify the extract constituents while 1D and 2D NMR spectroscopy were used for kojic acid (KA) structure confirmation. The antimicrobial, antioxidant, and cytotoxic activities were assessed in vitro. Both the extract and kojic acid showed potent antibacterial activity against Staphylococcus aureus and Pseudomonas aeruginosa with MIC 250 ± 0.82 µg/mL. Interestingly, the extract showed strong antifungal activity against Candida albicans and Cryptococcus neoformans with MIC 93.75 ± 0.55 and 19.53 ± 0.48 µg/mL, respectively. Furthermore, KA showed the same potency against Fusarium oxysporum and Cryptococcus neoformans with MIC 39.06 ± 0.85 and 39.06 ± 0.98 µg/mL, respectively. Ultimately, KA showed strong antioxidant activity with IC₅₀ 33.7 ± 0.8 µg/mL. Moreover, the extract and KA showed strong cytotoxic activity against colon carcinoma (with IC₅₀ 22.6 ± 0.8 and 23.4 ± 1.4 µg/mL, respectively) and human larynx carcinoma (with equal IC₅₀ 30.8 ± 1.3 and ± 2.1 µg/mL, respectively), respectively. The current study represents the first insights into the phytochemical profile and biological properties of P. chrysoenum ethyl acetate extract, which could be a promising source of valuable secondary metabolites with potent biological potentials.     

Baseline sensitivity and efficacy of trifloxystrobin against Sclerotinia sclerotiorum

The ascomycete fungus Sclerotinia sclerotiorum is a devastating plant pathogen notorious for its extremely broad host range. Trifloxystrobin has not been registered for control of S. sclerotiorum in China. In this study, baseline sensitivity of trifloxystrobin was established based on frequency distribution of 166 isolates' EC₅₀ values and efficacy of trifloxystrobin was determined on potted oilseed rape plants. Trifloxystrobin EC₅₀ values of the 166 isolates ranged from 0.01 to 0.80 μg/mL, with a mean value of 0.06 μg/mL. The frequency distribution of trifloxystrobin EC₅₀ values was unimodal in shape, but with a long right-hand tail. After logarithmic transformation, the frequency distribution fitted closer to a normal distribution than did the original EC₅₀ values. The preventive efficacies of trifloxystrobin at 5, 15, and 45 μg/mL were 71.4%, 96.5%, and 100.0%, respectively, while the curative efficacies were 40.6%, 48.7%, and 73.4%, respectively. Both preventive and curative efficacies of trifloxystrobin at 45 μg/mL were significantly higher (P ≤ 0.046) than those of the reference fungicide carbendazim. Assays with six arbitrarily selected isolates demonstrated that salicylhydroxamic acid (SHAM) at 20 μg/mL reduced trifloxystrobin EC₅₀ values by 84.0% on average and greatly potentiated efficacy of trifloxystrobin. The average co-toxicity factor of SHAM and trifloxystrobin for preventive efficacy was 64.3, indicating considerable synergisms in planta. Hence, SHAM should not be included in in vitro assay of S. sclerotiorum sensitivity to trifloxystrobin.     

Activities of essential oils from Asarum heterotropoides var. mandshuricum against five phytopathogens

Some secondary metabolites of plants function as antimicrobial products against phytopathogens and constitute an increasingly important class of pesticides. In the present study, the essential oil of Asarum heterotropoides var. mandshuricum was analyzed by GC/MS and its antimicrobial activity was evaluated against five phytopathogenic fungi. Major components of the oil were methyleugenol (59.42%), eucarvone (24.10%), 5-allyl-1,2,3-trimethoxybenzene (5.72%), and 3,7,7-trimethylbicyclo(4.1.0)hept-3-ene (4.93%). The essential oil and the most abundant component, methyleugenol, were separately assayed for inhibition of 5 pathogens: Alternaria humicola, Colletotrichum gloeosporioides, Rhizoctonia solani, Phytophthora cactorum and Fusarium solani. Both the oil and methyleugenol strongly inhibited the growth of the test pathogens (IC₅₀ values <0.42 μg ml⁻¹) except F. solani, with the best activity against P. cactorum (IC₅₀ values = 0.073 and 0.052 μg ml⁻¹, respectively). It is concluded that the essential oil of A. heterotropoides var. mandshuricum has a broad antiphytopathogenic spectrum, and that methyleugenol is largely responsible for the bioactivity of the oil. The mode of action of methyleugenol against P. cactorum is discussed based on changes in the mycelial ultrastructure.     

Use of essential oil of Laurus nobilis obtained by means of a supercritical carbon dioxide technique against post harvest spoilage fungi

The aspects of the antifungal activity of essential oil of laurel (Laurus nobilis) obtained by means of a supercritical carbon dioxide (SFE-CO₂) technique against post harvest spoilage fungi, have been studied in this research work by tests performed under in vitro and in vivo conditions. The measurement of antifungal activity of the oil, for its potential application as botanical fungicide, is very useful to find alternatives to synthetic fungicides. The present paper reports, for the first time, the results about the antifungal activity of laurel oil, obtained by a semi-industrial process that utilize a SFE-CO₂ technique, against three plant pathogenic fungi. The determination of the main active substances was carried out by gas chromatography analysis: laurel oil was characterized by high content (≥10%) of 1.8-cineole, linalool, terpineol acetate, methyl eugenol and a low content (<10%) of linalyl acetate, eugenol, sabinene, β-pinene, α-terpineol. The inhibition of the mycelial growth of Botrytis cinerea, Monilinia laxa and Penicillium digitatum was evaluated in vitro at the concentration range of 200, 400, 600, 800 and 1000 μg/mL. M. laxa was totally inhibited by application of the oil at the lowest concentration, B. cinerea was completely inhibited at the highest concentration, and a fungistatic action was observed in both cases. P. digitatum was only partially inhibited at all the concentration ranges. The activity of the oil, placed in the form of spray on the fruit skin at the concentration range of 1, 2 and 3 mg/mL, was studied by biological tests. Both curative and protective activities of the oil have been evaluated on peaches, kiwifruits, oranges and lemons artificially inoculated with M. laxa, B. cinerea and P. digitatum, respectively. A very good antifungal activity has been found on kiwifruits and peaches when the oil was placed before the inoculation at a concentration of 3 mg/mL (68 and 91% of decay inhibition respectively). The same activity has been found on peaches when the oil was placed after the infection (76% of decay inhibition). The application of the oil did not caused any phytotoxic effect and kept any fruit flavour, fragrance or taste. This study has demonstrated that the essential oil of L. nobilis extracted by a SFE-CO₂ technique, is one potential and promising antifungal agent which could be used as botanical fungicide in the postharvest protection of peaches and kiwifruits against M. laxa and B. cinerea.     

Different Culture Metabolites of the Red Sea Fungus Fusarium equiseti Optimize the Inhibition of Hepatitis C Virus NS3/4A Protease (HCV PR)

The endophytic fungus Fusarium equiseti was isolated from the brown alga Padina pavonica, collected from the Red Sea. The fungus was identified by its morphology and 18S rDNA. Cultivation of this fungal strain in biomalt-peptone medium led to isolation of 12 known metabolites of diketopeprazines and anthraquinones. The organic extract and isolated compounds were screened for their inhibition of hepatitis C virus NS3/4A protease (HCV PR). As a result, the fungal metabolites showed inhibition of HCV protease (IC₅₀ from 19 to 77 μM), and the fungus was subjected to culture on Czapek’s (Cz) media, with a yield of nine metabolites with potent HCV protease inhibition ranging from IC₅₀ 10 to 37 μM. The Cz culture extract exhibited high-level inhibition of HCV protease (IC₅₀ 27.6 μg/mL) compared to the biomalt culture extract (IC₅₀ 56 μg/mL), and the most potent HCV PR isolated compound (Griseoxanthone C, IC₅₀ 19.8 μM) from the bio-malt culture extract showed less of an inhibitory effect compared to isolated ω-hydroxyemodin (IC₅₀ 10.7 μM) from the optimized Cz culture extract. Both HCV PR active inhibitors ω-hydroxyemodin and griseoxanthone C were considered as the lowest selective safe constituents against Trypsin inhibitory effect with IC₅₀ 48.5 and 51.3 μM, respectively.     

Isolation, identification and dyeing studies of betanin on modified acrylic fabrics

Mature red fruits of Opuntia ficus-indica contain two soluble pigment, betanin and indicaxanthin. The optimal conditions for dye extraction were to mix 50 g of juice from cactus pears with 100 mL of acidified water as solvent for dye extraction. Two main dyes were purified from the pigment extract by chromatography and identified by UV-vis, HPLC and LC-MS techniques as indicaxanthin (15 mg per 100 g) and betanin (280 mg per 100 g). The effect of dye bath pH, salt concentration, dyeing time and temperature was studied. The optimal conditions for dyeing modified acrylic fabrics with betanin dye were carried out at 50 °C for 45 min at pH 5. Un-mordanted samples have good properties of water and washing fastness. Mordant CoSO₄ was found to give good light fastness (rating 5).     

Screening and quantification of an antiseptic alkylamide, spilanthol from in vitro cell and tissue cultures of Spilanthes acmella Murr.

The study revealed, for the first time, accumulation of spilanthol, an antiseptic alkylamide, in in vitro cultures of Spilanthes acmella Murr., a medicinal plant of immense commercial value. To achieve this, in vitro shoots were regenerated via direct organogenesis from leaf-disc explants of Spilanthes. Shoots were induced in the presence of N⁶-benzylaminopurine (BAP) alone or in combination with either α-naphthalene acetic acid (NAA) or Indole-3-acetic acid (IAA) in Murashige and Skoog medium. The best treatment for shoot regeneration was MS + BAP (5.0 μM) + IAA (5.0 μM), which promoted adventitious shoot proliferation in >82% cultures with an average of 5.3 shoots per explant. Regenerated shoots rooted spontaneously with a frequency of 100% on half strength MS medium (major salts reduced to half strength) containing 50 g l⁻¹ sucrose. The plantlets were acclimatized successfully with 90% survival rate. Additionally, ploidy stability of the regenerated plants was assessed by flow cytometry which showed that all investigated plants had the similar ploidy as that of the mother plant. For spilanthol identification, peaks eluted from HPLC were analyzed by mass spectrometry with its characteristic fragmentation pattern. For quantification studies, calibration curve was generated, which revealed a higher amount of spilanthol content (3294.36 ± 12.4 μg/g DW) in the leaves of in vitro plants compare to those of in vivo plants (2703.66 ± 9.6 μg/g DW of spilanthol). An efficient multiplication frequency, ploidy stability and enhanced spilanthol accumulation ensure the efficacy of the protocol developed for this industrially important medicinal plant.     

Identification and antimicrobial activity of two alkaloids from traditional Chinese medicinal plant Tinospora capillipes

Two antimicrobial alkaloids, palmatine and jatrorrhizine, were isolated from tubers of traditional Chinese medicinal plant Tinospora capillipes using activity-guided isolation method and chromatography. Their antimicrobial activity was determined in vitro. The results showed that palmatine and jatrorrhizine had inhibitory activity against plant pathogens Colletotrichum gloeosporioides, Fusarium oxysporum f. sp. niveum, Mycosphaerella sentina, Pestalotia mangiferae, Cercospora kaki, Gymnosporangium haraeanum, Rhizoctonia solani and Colletotrichum graminicola, with the EC₅₀ values of 0.0348-0.8356 g L⁻¹ and 0.0240-0.8649 g L⁻¹, respectively. Palmatine and jatrorrhizine also exhibited inhibition against animal pathogens Bacillus cereus, Bacillus megaterium, Bacillus subtilis, Staphyloccocus aureus, Staphylococcus epidermidi, Micrococcus lysodeikticus, Proteus vulgaris, Salmonella typhi and Escherichia coli, with the MIC values of 0.1-0.8 g L⁻¹ and 0.1-0.6 g L⁻¹, respectively. These results suggested that palmatine and jatrorrhizine showed relatively broad spectrum antimicrobial activity against plant and animal pathogens.