Ethanolic extract of mango (Mangifera indica L.) malformed inflorescence as effective antifungal agent

Mango (Mangifera indica L.) suffers from floral and vegetative malformation and crop production is seriously affected. The anti-fungal activity of ethanolic extract of malformed mango inflorescence was observed at different concentrations (1000, 2000, 3000, 4000 and 5000 μl/ml) against 10 fungi, viz., Ustilago cynodontis, Cercospora cajani, Sphaerotheca sp., Cercospora sp., Alternaria solani, Bipolaris sp., Helminthosporium sp., Curvularia sp., Fusarium udum and Alternaria cajani. Spore germination of most of the fungi was inhibited at 5000 μg/ml. Some of them were also susceptible at 3000 or 4000 μg/ml concentration. Analysis of phenolic acids by high performance liquid chromatography (HPLC) showed 18 peaks in the extract, but only four could be identified, viz., capachin, gallic, benzoic and cinnamic acids. Because of the high efficacy of ethanolic extract of malformed mango inflorescence, its use under field conditions to control some plant diseases has been suggested.     

Antifungal efficacy of some ethyl acetate extract fractions of Cyperus rotundus rhizomes against spore germination of some fungi

Plant products play an important role as safe and ecofriendly method in controlling various plant diseases. Ethyl acetate fractions of Cyperus rotundus rhizomes have been found highly effective against some species of Alternaria (A. alternata, A. brassicola, A. solani, Alternaria chearanthi), Colletotrichum (C. musae, Colletotrichum sp.), Curvularia (C. lunata, C. maculans, C. pallescens, C. pennisetti), Helminthosporium (H. pennissetti, H. spiciferum, H. echinoclova and Heterosporium colocasiae) in vitro. A. brassisicola was highly sensitive to all the fractions at all the concentrations. Fractions 4–7 inhibited complete spore germination at 2000 and 3000 μg/ml as compared to control. However, 100% spore germination inhibition was found in Colletotrichum species in all the fractions at 2000 and 3000 μg/ml but at lower concentrations 50–60% spore germination and 90% reduction of germ tube elongation were observed. Curvularia species was highly sensitive to all concentrations of all the fractions as 50–100% spore germination inhibition was recorded at 500 μg/ml dose. Some species of Helminthosporium were sensitive at 3000 μg/ml in some fractions but other test fractions showed least efficacy. Fractions 4–7 were 100% inhibitory for H. colocasiae. Germ tube elongation was also affected by 60–90%. The germ tube branching and their elongation were affected in almost all species at 30 to 95%. The high efficacy of ethyl acetate fractions of rhizomes of C. rotundus against some fungi indicates that they can be very well tried under field conditions against some important plant diseases as an ecofriendly method of plant disease control.     

Antifungal and high-performance liquid chromatography analysis of the crude extracts of Acorus calamus,Tinospora cordifolia and Celestrus paniculatus

The antifungal activity of methanolic crude extracts of Acorus calamus, Tinospora cordifolia and Celestrus paniculatus was investigated against Alternaria solani, Curvularia lunata, Fusarium sp., Bipolaris sp. and Helminthosporium sp. at different concentrations (1000, 2000, 3000, 4000 and 5000 μg/ml). At 5000 μg/ml, crude extract of T. cordifolia is found to be highly effective against Helminthosporium sp. followed by A. calamus against A. solani. On the other hand, at 5000 μg/ml, C. paniculatus showed better activity against A. solani and Helminthosporium followed by A. calamus against A. solani at 4000 μg/ml. At 5000 μg/ml, all the three crude extracts showed least activity against fungus C. lunata and Fusarium sp. except A. calamus that showed better activity against C. lunata. The increase in the production of phenolic acid in the extract can be correlated with the induction of resistance in treated plants against phytopathogenic fungi. High-performance liquid chromatography analysis of the crude extract of medicinal plants showed six different phenolic acids (benzoic acid, cinnamic acid, caffeic acid, ferulic acid, gallic acid and tannic acid) present in varying amounts. The results of the study provide scientific basis for the use of the plant extract in the future development as antioxidant, antibacterial, antifungal and anti-inflammatory agent.     

Dieter Seidel‐65 Jahre

Fruit extract of Solanum xanthocarpum was evaluated for its toxicity against Alternaria brassicae, the causal agent of Alternaria blight of Indian mustard [Brassica juncea (L.) Czern. &; Coss]. The mass obtained after vacuum drying of the crude methanolic extract was utilised for further sequential fractionation using n-hexane, ethyl acetate, n-butanol and methanol. Among the crude and different fractions tested, methanolic fraction was most effective with a minimum inhibitory concentration (MIC) of 62.5 μg/ml. The methanolic fraction was further fractionated using open column liquid chromatography into five subfractions (I–V) to identify the antifungal bioactive compounds. Among the five subfractions (SFs) tested SF IV was most effective at inhibiting A. brassicae conidial germination and thereby inhibited lesion development of Alternaria blight at a concentration of 15.625 μg/ml or higher. Furthermore, bioautography of SF IV with Alternaria alternata and diagnosis with Dragendorff reagent indicated that SF IV contains a mixture of bioactive alkaloids, namely a₁ (R_f = 0.12) and a₂ (R_f = 0.22). The potential of using S. xanthocarpum as a resource for the development of biofungicides is discussed.     

Effect of copper ions on the ligninolytic enzyme complex and the antioxidant enzyme activity in the white-rot fungus Trametes trogii 46

White-rot fungi of the Phylum Basidiomycota are quite promising in ligninolytic enzyme production and the optimization of their synthesis is of particular significance. The aim of this study was to investigate the effect of enhanced concentration of copper (Cu) ions (25–1000 μg/ml) on the activity of the ligninolytic enzyme complex (laccase, Lac; lignin peroxidase, LiP; Mn-peroxidase, MnP) in Trametes trogii 4₆, as well as the changes in the antioxidant cell response. All concentrations tested reduced significantly in growth and glucose consumption. Cu ions affected the ligninolytic enzyme activity in a dose dependent manner. Concentrations in the range of 25–100 μg/ml strongly stimulated Lac production (a 5–6-fold increase compared to the control). LiP activity was also induced by Cu, with the peak value being recorded following exposure to 50 μg/ml metal ions. In contrast, the addition of Cu ions had a positive effect on MnP activity at a concentration higher than 100 μg/ml. The maximum enzyme level was achieved at 1000 μg/ml. The results obtained on superoxide dismutase and catalase activities indicated that exposure of T. trogii 4₆ mycelia to Cu ions promoted oxidative stress. Both enzyme activities were co-ordinately produced with Lac and LiP but not co-ordinately with MnP.     

Catesbeianin-1, a novel antimicrobial peptide isolated from the skin of <Emphasis Type="Italic">Lithobates catesbeianus</Emphasis> (American bullfrog)

Objectives

To identify and characterize a novel antimicrobial peptide, catesbeianin-1.

Results

Catesbeianin-1 is 25 amino acids long and is α-helical, cationic and amphipathic. It had antimicrobial activity against Gram-positive and Gram-negative bacteria. It was resistant against trypsin and pepsin. Catesbeianin-1 exhibited moderate hemolytic activity (approx 8%) at 100 μg/ml, and its HC₅₀ (50% hemolytic concentration) was 300 μg/ml. Its cytotoxicity was approx 10–20% at 100 μg/ml, and its CC₅₀ (50% cytotoxic concentration) was >100 μg/ml. The LD₅₀ of catesbeianin-1 in mice was 80 mg/kg. At 3.1 µg/ml, catesbeianin-1 significantly inhibited the growth of methicillin-resistant Staphylococcus aureus.

Conclusions

A new antimicrobial peptide from the skin of Lithobates catesbeianus (American bullfrog) may represent a template for the development of novel antimicrobial agents.

     

Effect of plant secondary metabolites on common cutworm,Spodoptera litura (Lepidoptera: Noctuidae)

The effect of various flavonoids, lectins and phenyl β‐_D‐glucoside on larval survival, weights and the activities of digestive (total serine protease and trypsin) and detoxifying (esterase and glutathione‐S‐transferase) enzymes of Spodoptera litura larvae at 7 days after treatment was studied through diet incorporation assay. Flavonoids (rutin, chlorogenic acid, quinic acid, caffeic acid, naringenin, quercitin, kaempferol, myricetin, catechin, and ferulic acid) were incorporated in artificial diet at 100, 500 and 1000 ppm, lectins: groundnut leaf lectin (GLL), concavalin A (ConA) and phenyl β‐_D‐glucoside at 1, 2 and 5 μg/mL. Flavonoids such as rutin, quercitin and kaempferol at 1000 ppm were more toxic to S. litura larvae than quinic acid, caffeic acid, naringenin, myricetin, catechin, and ferulic acid. Larval growth and development were significantly reduced in S. litura larvae fed on a diet with GLL and ConA at 5 μg/mL compared to the larvae fed at 2 and 1 μg/mL concentrations. The larvae fed on flavonoid‐treated diets showed significant reduction in serine protease, trypsin and esterase activities. The flavonoids such as rutin, chlorogenic acid, quinic acid, naringenin, quercitin, kaempferol and myricetin, and lectins, GLL and ConA can be utilized in insect control programs.     

Antioxidant and Neuroprotective Effects of Scrophularia striata Extract Against Oxidative Stress-Induced Neurotoxicity

In this study, the neuroprotective effect of Scrophularia striata Boiss (Scrophulariaceae) extract, a plant growing in northeastern of Iran, against oxidative stress-induced neurocytotoxicity in PC12 was evaluated. The PC12 cell line pretreated with different concentrations (10, 50, 100, and 200 μg/ml) of the extract and then treated with H₂O₂ to induce oxidative stress and neurotoxicity. Survival of the cells, reactive oxygen species (ROS) generation, and apoptosis were measured using MTT assay, fluorescent probe 2′,7′-dichlorofluorescein diacetate, and annexin V/propidium iodide, respectively. Moreover, the 2,2-diphenyl-1-picryl-hydrazyl (DPPH) was used to evaluate the antioxidant capacity of the plant extract. Phytochemical assay by thin layer chromatography showed that the main components, including phenolic compounds, phenyl propanoids and flavonoids, were presented in the S. striata extract. The extract in concentrations of 50–200 μg/ml protected PC12 cells from H₂O₂-induced toxicity. The survival of the cells at concentration of 200 μg/ml was 64 % compared to that of H₂O₂ alone-treated cells (48 %) (p < 0.001). The extract also dose-dependently reduced intracellular ROS production (p < 0.001). Moreover, the extract showed antioxidative effects and decreased apoptotic cells. Collectively, these findings indicated the ability of S. striata to decrease ROS generation and cell apoptosis and also suggest the presence of the neuroprotective agents in this plant.     

Antifungal activity of two alkaloids of Zephyranthes citrina and their field efficacy against powdery mildew (Erysiphe cichoracearum) of balsam

The anti-fungal activity of two alkaloids isolated from bulbs of Zephyranthes citrina was observed against 10 fungi, viz, Aternaria solani, A. triticina, Curvularia lunata, C. maculuns, Cercospora malvacearum, Erysiphe sp., Fusarium udum, Helminthosporium pisi, H. speciferum and Ustilago cynodontis. Different concentrations (200, 400, 600, 800, 1000 µg/ml of alkaloids (A and B) were used. Spore germination was inhibited at 600, 800, 1000 µg/ml. B alkaloid was used against Erysiphe cichoracearum causing powdery mildew in balsam (Impatiens balsamina) in the field as pre- and post-inoculation treatments at 1000, 1500, 2000 µg/ml doses. The extract was effective in both pre- and post-inoculation treatments. Foliar application of this alkaloid resulted in inducing synthesis of phenolic acid in the leaves of balsam. Maximum phenolics were detected in the leaves treated with 1500 µg/ml in both pre- and post-inoculation treatments. The increase in the production of phenolics in treated leaves of balsam can be correlated with the induction of resistance in treated plants against powdery mildew. The significant efficacy of the alkaloid under field conditions opens the possibility of its use by farmers for also controlling other diseases.     

In vitro and in silico characterization of angiogenic inhibitors from <Emphasis Type="Italic">Sophora interrupta</Emphasis>

Sophora interrupta Bedd, (Fabaceae) is used in Indian folk medicine to treat cancer. Angiogenesis is one of the crucial characteristics of cancer metastasis and is regulated by vascular endothelial growth factor (VEGF). In this study, we examined the antiangiogenic properties of the root ethyl acetate extract of Sophora interrupta by various methods. In vitro antioxidant activity (100–600 μg/ml) of S. interrupta ethyl acetate (SEA) extract was evaluated by DPPH and ABTS, anti-inflammatory activity (50, 100 and 150 μg/ml) by estimating nitric oxide (NO) levels, anti-angiogenic activity (200 and 500 μg/ml) was validated by chorio allantoic membrane (CAM) assay and in silico molecular dynamic (MD) simulations analyses (25 ns) were performed to identify the anti-angiogenic compounds extracted from root extract. The antioxidative activity of SEA extract at IC₅₀ (200?±?0.6 μg/mL) is equal to that of ascorbic acid at IC₅₀ (50?±?0.6 μg/mL), and the anti-inflammatory activity of SEA extract at IC₅₀ (150?±?0.2 μg/mL) was inhibited significantly by nitric oxide (NO) production. The SEA extract significantly reduced the sprouting of new blood vessels at ID₅₀ 500?±?0.13 μg/mL in the CAM assay. Gas chromatography–mass spectrometry analysis of the SEA extract detected 34 secondary metabolites, of which 6a,12a-dihydro-6H-(1,3)dioxolo(5,6)benzofuro(3,2-c)chromen-3-ol (maackiain) and funiculosin formed strong hydrogen bond interactions with Lys 920, Thr 916 and Cys 919 (2H), as well as Glu 917 of VEGFR2, and these interactions were similar to those of the anti-angiogenic compound axitinib. Significant findings in all the assays performed indicate that SEA extract has potential anti-angiogenic compounds that may interfere with VEGF-induced cancer malignancy.     

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca inhibits spore germination of some fungi

The mixture of tertiary and quaternary alkaloids isolated from Argemone ochroleuca was separately assessed against spore germination of some plant pathogenic fungi, e.g. Alternaria alternata, Alternaria brassicae, Alternaria cajani, Bipolaris sp., Curvularia lunata, Curvularia sp., Colletotrichum musae, Fusarium udum, Helminthosporium sp., Helminthosporium pennisetti and Helminthosporium speciferum. Spore germination of Fusarium udum and Helminthosporium sp. was completely inhibited at very low concentration (200 ppm). A similar effect was observed on A. alternata, C. musae and H. pennisetti at 600, 800 and 1000 ppm. With quaternary alkaloids, Curvularia sp. and Colletotrichum musae were most sensitive as complete inhibition of spore germination was observed at 400, 600, 800 and 1000 ppm and a similar effect was observed with A. brassicae and A. cajani at 600, 800 and 1000 ppm. The remaining fungi were also highly sensitive to the mixture at different concentrations.     

Beneficial effects of a medicinal herb,Cirsium japonicum var. maackii,extract and its major component,cirsimaritin on breast cancer metastasis in MDA-MB-231 breast cancer cells

The biological activities of the ethanol extract from Cirsium japonicum var. maackii (ICF-1) and its major component, polyphenol cirsimaritin, were investigated as part of the search for possible alternative drugs for breast cancer. Three in vitro cell-based assays were used: the cell proliferation assay, tube-formation assay, and Western blot analysis. Both the ICF-1 extract and cirsimaritin inhibited the viability of HUVECs in a dose-dependent manner. The inhibition achieved was 36.89% at a level of 200 μg/ml by the ICF-1 extract and 62.04% at a level of 100 μM by cirsimaritin. The ICF-1 extract and cirsimaritin reduced tube formation by 12.69% at level of 25 μg/ml and 32.18% at the levels of 6.25 μM, respectively. Cirsimaritin inhibited angiogenesis by downregulation of VEGF, p-Akt and p-ERK in MDA-MB-231 cells, suggesting that cirsimaritin is potentially useful as an anti-metastatic agent. The present study demonstrated that Cirsium japonicum extract and its active component cirsimaritin is an excellent candidate as an alternative anti-breast cancer drug.     

Anti-herpesvirus bovine type 5 activities of extracts obtained from Plocamium brasiliense

Bovine herpesvirus type 5 (BoHV-5) is an important etiologic agent of meningoencephalitis in cattle and has been frequently identified in outbreaks of neurological disease in bovine in the southern hemisphere including Brazil. This study aimed to evaluate the cytotoxic effect and the antiviral properties of extracts obtained from Plocamium brasiliense (Greville) Howe and Taylor in BoHV-5 RJ42/01 replication. The cytotoxic effects were measured in Madin-Darbin bovine kidney cells (MDBK) and cytotoxic concentration (CC₅₀) values have been determined for acyclovir (ACV) (223 μg?±?2.0), ethyl acetate extract from P. brasiliense (2,109 μg?±?10), hexane extract from P. brasiliense (7.181 μg?±?5), dichloromethane extract from P. brasiliense (2.356 μg?±?6.5), and hydroalcoholic extract from P. brasiliense (1.408 μg?±?5.8). As a first approach to characterize the action of these extracts on BoHV-5 RJ42/01, a virucidal assay activity was performed. A virus suspension containing 1?×?10⁵ plaque-forming units (PFU) of the BoHV-5 RJ42/01 was mixed with 600 μg of extract and acyclovir and kept at room temperature (24 °C) for 3 h. Meanwhile, a control of untreated infected virus was performed in the same conditions. Then, treated virus suspension and untreated control were diluted, and percentage of inhibition of infectivity was determined by plaque assay: ethyl acetate extract (99 %), hexane extract (90 %), dichloromethane extract (99 %), and hydroalcoholic extract (27 %). Acyclovir had a slight virucidal activity on viral particle. The inhibition of attachment was performed in MDBK cells inoculated with 100 PFU of BoHV-5 RJ42/01 in the presence or absence of various concentrations of extracts (0.3, 0.9, and 1.5 μg mL^?1). Acyclovir was also assayed at 2.8 and 11.25 μg mL^?1. The inhibition of adsorption was also tested in MDBK cells treated with the same concentrations of the extracts before virus inoculation. Results: hexane extracts inhibited virus attachment in pre-treated cell 0.9 μg (55 %) and 1.5 μg (71 %) and untreated MDBK cell only with 1.5 μg (63 %). Ethyl acetate extract on cell pre-treated with 0.3 μg (67 %), 0.9 μg (81 %), and 1.5 μg (91 %). Ethyl acetate extract on pre-treated cell 0.3 μg (67 %), 0.9 μg (81 %), and 1.5 (91 %) but discrete inhibition on cell untreated. Dichloromethane extract and acyclovir slightly inhibited virus binding on MDBK cell.     

Isolation and identification of secondary metabolites from hexane extract of culture filtrate of Bacillus licheniformis MTCC 7445

Hexane extract of cell free culture filtrate of Bacillus licheniformis MTCC 7445 isolated from rhizoshere soil of a resistant tomato plant showed antifungal activity against a number of soilborne and human pathogenic fungi. Maximum activity was observed against Botrytis cinerea (ED₅₀ = 23.79 μg/ml), Candida albicans (ED50 33.45 μg/ml) and Microsporum canis (ED₅₀ = 39.02 μg/ml). Metabolites such as 1-methyl pyrrolidene, 1-methyl cyclohexene, 4,4-dimethyl cyclohexane, ethyl-4-ethoxybenzoate, 2-butoxyethanol, naphthalene, ter butyl benzene and phenoxy acetic acid were identified by GC-MS and comparing the mass spectrum with the NIST library.     

A polyphenolic compound rottlerin demonstrates significant in vitro cytotoxicity against human cancer cell lines: isolation and characterization from the fruits of Mallotus philippinensis

In vitro assay for cytotoxic activity of glands/hairs obtained from the fruits of Mallotus philippinensis has been carried out against 14 human cancer cell lines from nine different origins via 95% ethanolic, 50% ethanolic and aqueous extract at the concentration of 100 μg/ml. Results revealed that the 95% ethanolic extract showed highest in vitro cytotoxic effect against all the 14 human cancer cell lines. The fractions of the same extract i.e. 95% ethanolic were obtained and it was found that the significant cytotoxic potential was produced by the chloroform soluble fraction at 100 μg/ml as this fraction inhibited the growth of ten human cancer cell lines from seven different tissues. Further, the chromatographic analysis of the said fraction afforded a polyphenolic molecule rottlerin. This drug at the concentration of 1?×?10^?5M and 1?×?10^?4M suppressed the proliferation of eight human cancer cell lines from six different tissues and proved its exceptionally remarkable in vitro anticancer efficiency.     

Effect of carbendazim and mancozeb combinationon Alternaria leaf blight and seed yield in sunflower (Helianthus annus L.)

Abstract

Leaf blight caused by Alternaria helianthi (Hansf.) Tubaki & Nishihara, is the major disease of sunflower affecting the successful cultivation across India. Five individual fungicides and two combination fungicides were evaluated against this pathogen in laboratory and in field experiments. Among them, the combination of carbendazim + mancozeb completely (100%) inhibited the mycelial growth of A. helianthi, irrespective of the concentrations tested followed by carbendazim alone and metalaxyl + mancozeb under in vitro condition. In field conditions, the combination of carbendazim + mancozeb was found to be highly effective in reducing the leaf blight disease of sunflower in all the three experiments as compared to other fungicides and unsprayed control. The reduction of Alternaria leaf blight was also directly associated with an increase in seed yield. The economics of the fungicides spray has been worked out and the benefit cost ratio for the combination of carbendazim + mancozeb at 2.0 g/l was 7.1 as compared to unsprayed control. The overall analysis of the results revealed that the combination of carbendazim + mancozeb at 2.0 g/l can be used for the management of foliar diseases such as Alternaria leaf spot/blight in agricultural crops.     

Variation of loganin content in Cornus officinalis fruits at different extraction conditions and maturation stages

Efficient preparation of loganin from Cornus officinalis fruits was investigated. First, effect of extraction conditions on loganin yield was measured. The loganin content in C. officinalis extract was greatly affected by ethanol concentration and extraction time whereas extraction temperature exerted relatively little effect. Response surface methodology with Box–Behnken design suggested optimized extraction condition for maximum loganin yield as ethanol concentration, 32.0%; temperature 46.2 °C and extraction time, 46.7 min, which yielded 10.4 μg loganin/mg dried fruit. Next, the effect of maturation stage of C. officinalis fruits on loganin content was investigated. The loganin content in the extract of C. officinalis fruits was decreased as the maturation process. The loganin content in the unripe fruits was 18.0 μg/mg extract whereas reduced to 13.3 μg/mg extract for ripe fruits. Taken together, our present study suggested the importance of extraction condition and maturation stages for efficient preparation of loganin from C. officinalis fruits.     

Variation of Fungicide Resistance in Czech Populations of Pseudoperonospora cubensis

During the growing seasons between the years 2001 and 2004, 98 isolates of Pseudoperonospora cubensis from nine regions of Czech Republic were collected and screened for tolerance/resistance to the three frequently used fungicides (propamocarb, fosetyl‐Al, metalaxyl). Fungicides were tested in five different concentrations, using a floating disc bioassay. Fungicide effectiveness varied considerably. Propamocarb appeared most effective and all the isolates collected in the years 2001–2003 were found sensitive to all tested concentrations [607–9712 μg active ingredient (a.i.)/ml]. In 2004, some strains with increased resistance to propamocarb were detected. These strains were characterized by tolerance at the lowest concentrations (607 μg a.i./ml, eventually on 1214 μg a.i./ml); however, they were controlled by 2428 μg a.i./ml. Fosetyl‐Al was effective at the recommended concentration of 1600 μg a.i./ml against all isolates. However, the occurrence of isolates (collected in 2001) which sporulated at low concentrations (400 and 800 μg a.i./ml) indicated that the selection for tolerance occurs in the pathogen population. Nevertheless, this phenomenon was not confirmed with the P. cubensis isolates collected between the years 2002 and 2004. Metalaxyl was found ineffective, because 97% of the isolates showed the resistance to the recommended concentration (200 μg a.i./ml), and the other 3% of isolates expressed tolerant response. The majority of the isolates showed profuse and/or limited sporulation at higher concentrations (400 and 800 μg a.i./ml). A substantial shift to highly metalaxyl resistant strains was evident in the Czech P. cubensis populations during 2001–2004.     

Repellency and toxicity of extract from Francoeria crispa (Forsk.) to Eutetranychus orientalis (Klein) (Acari: Tetranychidae)

Laboratory bioassays were conducted to characterise the activity of the extract Francoeria crispa (Forsk.) (Family: Compositae) against the citrus brown mite, Eutetranychus orientalis (Klein). Ethyl acetate was tested for preparing the crude extract of F. crispa. The extract was tested for its toxicity against eggs and adult females of the pest E. orientalis. Ethyl acetate extract of F. crispa affected the behaviour, toxicity and fecundity of females under laboratory conditions. The extract had similar toxic effects on egg stage and adult females of E. orientalis (LC₅₀ = 0.00050 g/ml), respectively. Leaf discs treated with increasing concentrations of ethyl acetate extract of F. crispa showed a high percentage of repellency (97.45%), respectively. Treated females with LC₅₀ concentration of ethyl acetate extract showed a higher remarkable percentage of mortality as well as a reduction in the total number of eggs laid during 7 days. Ten isolated fractions of ethyl acetate crude extract from F. crispa were detected. The results clearly indicate that the isolate number (10) was the most toxic isolate on eggs and females of E. orientalis (LC₅₀ = 0.00014 and 0.000125 g/ml), respectively.     

Differences in the susceptibility of five herbivore species and developmental stages to tomato resistance induced by methyl jasmonate treatment

We compared the susceptibility of five herbivores to tomato resistance induced by methyl jasmonate (MeJA) treatment. We tested for lethal effects against five herbivores (Spodoptera litura, Mamestra brassicae, Frankliniella occidentalis, Tetranychus urticae, and Henosepilachna vigintioctopunctata) at various MeJA concentrations. The mortality of all five herbivores increased significantly with increasing MeJA concentration. The 25 % lethal concentration was 0.03 μM for both first-instar larvae of S. litura and third-instar larvae of M. brassicae, 0.51 μM for third-instar larvae of S. litura, 0.76 μM for adult T. urticae, 2.4 μM for first-instar larvae of F. occidentalis, and 5.7 μM for first-instar larvae of H. vigintioctopunctata. Thus, the degree of susceptibility to MeJA-induced resistance of tomato was first-instar larvae of S. litura = third-instar larvae of M. brassicae > third-instar larvae of S. litura ≈ adult T. urticae > first-instar larvae of F. occidentalis > first-instar larvae of H. vigintioctopunctata. Mortality of first-instar larvae of M. brassicae was >90 % at all concentrations. Mortality of fourth-instar larvae of H. vigintioctopunctata (<7 %) was similar to that of the control at all MeJA concentrations. We also detected statistically significant weight loss of the surviving lepidopteran larvae, increased larval duration of F. occidentalis and H. vigintioctopunctata, and reduced egg production by T. urticae grown on MeJA-treated tomato, suggesting that the MeJA-induced resistance can control these herbivores, but effectiveness is different on different species and growth stage. Feeding by both M. brassicae and H. vigintioctopunctata larvae activated JA-inducible genes in tomato.