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.     

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.     

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.     

Effect of boron on the development of brown rot (Monilinia laxa) on peaches

Requirements of consumers for products with low residues of pesticides have increased the need for alternative disease management practices. The concentration of boron in fruit affects its quality, shelf life and the development of physiological disorders. However, the effect of boron on the susceptibility of peach to fruit rots has not been reported. This study investigated the effect of boron (Power B and Borax) on the development of Monilinia laxa on peaches (cv Andross). Mycelial growth of M. laxa was inhibited on potato dextrose agar supplemented with 750 μg ml⁻¹ of Borax or 1000 μg ml⁻¹ of Power B. The EC 50 values were 107.9 and 522.4 for Borax and Power B respectively. Field investigations showed that the incidence of peach infections by M. laxa was negatively correlated with the content of Boron in the leaves. Post-harvest dipping of peaches in Power B or Borax solution, at concentrations recommended by manufacturer (2 μg ml⁻¹ for Power B and 1 mg ml⁻¹ for Borax), significantly reduced the development of M. laxa. Power B, at rates of 6 μg ml⁻¹, and Borax at rates of 3 mg ml⁻¹ were the most effective in reducing infections by M. laxa. Finally, post-harvest dipping of fruit in Power B or Borax reduced losses of fruit weight and improved fruit firmness one month after storage, showing that boron increased the maintainability of peaches in cold storage. Peaches treated with 6 μg ml⁻¹ Power B or 3 mg ml⁻¹ Borax had the highest flesh firmness and the lowest water losses, while untreated control peaches were the least firm. Generally, Borax was significantly less effective than Power B, but better than the control treatment.     

Potential of extracts of the tropical plant Balanites aegyptiaca (L) Del. (Balanitaceae) to control the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae)

The effects of extracts of different parts of the perennial tropical plant Balanites aegyptiaca (L) Del., including various solvent extracts of roots, methanol extracts from leaves, fruits, flowers and roots, partially purified saponins obtained from its roots and a standard saponin were studied on the life cycle (adult longevity, number of eggs, crawlers, adults, weight of adults and % wax content) of a laboratory-reared parthenogenic line of the mealy bug, Maconellicoccus hirsutus (Homoptera: Pseudococcidae). Extracts derived from various parts of B. aegyptiaca (leaves, fruits, flowers, and roots in methanol) affected the life cycle of M. hirsutus with a methanol root extract being the most effective at a concentration of 500 μg ml⁻¹. Partially purified saponin of B. aegyptiaca and the commercial bark saponin extract (Sigma) from Quillaja saponaria at a concentration of 500 μg ml⁻¹ were effective in reducing the longevity of M. hirsutus. Significant reductions in oviposition by M. hirsutus were found for all the extracts at a concentration of 500 μg ml⁻¹. Extracts also affected the number of emerging crawlers, number of adults as well as the weight and wax content of emerging adults. These studies suggest that B. aegyptiaca plant extracts and saponins can be useful botanical insecticides for the protection of crops from mealy bugs.     

Adventitious root cultures of Castilleja tenuiflora Benth. as a source of phenylethanoid glycosides

Castilleja tenuiflora is a highly valued medicinal plant that grows in pine-oak woods in Mexico. In this study, we identified for the first time verbascoside and isoverbascoside as the major phenylethanoid glycosides (PhGs) in C. tenuiflora. These compounds have proven biological activities, including anti-inflammatory, antioxidant, and cytotoxic activities, which may be related to the traditional uses of this plant. We developed a reverse-phase high-performance liquid chromatography (RP-HPLC) procedure to analyze PhGs, and determined their concentrations in various different tissues of wild plants. Verbascoside accumulated mainly in roots and inflorescences (9.23 and 7.88 mg g⁻¹ dry biomass, respectively), while isoverbascoside accumulated mainly in the roots (7.13 mg g⁻¹ dry biomass). To provide an alternative source of material for production of bioactive compounds, we established in vitro adventitious root cultures in which roots were grown in B5 medium containing either 10 μM indole 3-acetic acid (IAA) or 10 μM α-naphthaleneacetic acid (NAA). The greatest dry biomass yield (30 g L⁻¹) was achieved at 30 days after transfer of roots into IAA-containing medium. The highest specific yields of PhGs were also obtained using this auxin; the maximum level of verbascoside was 14.62 mg g⁻¹ dry root biomass (438.6 mg L⁻¹) at 30 days after root transfer, and the maximum yield of isoverbascoside was 37.32 mg g⁻¹ dry root biomass (522.48 mg L⁻¹) at 23 days after root transfer. Adventitious root cultures of C. tenuiflora are a promising system for further studies on scale-up and phenylethanoid glycosides biosynthesis.     

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.     

Insecticidal and repellent activity of selected essential oils against of the pollen beetle, Meligethes aeneus (Fabricius) adults

The essential oils from 9 aromatic plants were tested on repellency and mortality of Meligethes aeneus adults. All the tested essential oils caused high mortality of M. aeneus adults in the tarsal tests. The lethal doses after 6 h exposure were ranged between 197 and 1508 μg cm⁻². Essential oils obtained from Carum carvi and Thymus vulgaris were most efficient where LD₅₀ was estimated as 197 and 250 μg cm⁻², respectively.Repellency declined in all the essential oils as a function of time. The longest persistence time was determined for essences obtained from C. carvi and T. vulgaris where significantly the highest repellent index of 65.6% and 63.8%, respectively, was determined. Repellent index lower than 15% was determined for the remaining essential oils.     

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.     

Nematicidal efficacy of isothiocyanates against root-knot nematode Meloidogyne javanica in cucumber

Isothiocyanates (ITCs), a series of new nematicides of the -NCS group, were evaluated for their efficacy against root-knot nematode Meloidogyne javanica. Of the compounds tested, AllylITC, AcITC, EtITC, BzTC, BzITC, 1-PEITC and 2-PEITC showed in vitro irreversible nematicidal activity against second-stage juveniles of M. javanica, following exposure for 72 h at concentrations as low as 5 μg mL⁻¹. When exposed to AllylITC, AcITC and EtITC at lower concentrations, motile juveniles also became irreversibly immobile in 3 days, with a LC₅₀ value at 2.76, 2.53 and 3.05 μg mL⁻¹, respectively. In the pot experiments, 1.0 ml AllylITC and 1.1 ml AcITC per kg of soil controlled M. javanica, similarly to or better than metam sodium at its recommended dose. Similar results were obtained in the field experiments using 1.0 kg AllylITC or 1.0 kg AcITC ha⁻¹. Based on the results of this study, AllylITC and AcITC have potential to be used as new bio-fumigant nematicides.     

Highly efficient in vitro regeneration of the industrial oilseed crop Crambe abyssinica

A highly efficient regeneration protocol for oilseed crop Crambe abyssinica has been developed using hypocotyls as explants in this study. Crambe is a potential engineering oilseed crop for industrial purposes as it contains 55-60% erucic acid in its oil and, more importantly, it does not outcross with any food oil seed crops. However, the low regeneration frequency with the currently available protocols is still a limiting factor for genetic modification of Crambe. In this study, we investigated the effects of N-source, C-source, AgNO₃, cultural conditions as well as the concentration and combination of plant growth regulators (PGR) on the regeneration frequency of C. abyssinica. The results showed that all these factors, especially the N-source and PGR concentrations and combinations, played an important role in shoot regeneration. Among all the factors tested, the combination of using hypocotyls from C. abyssinica cv. galactica, the Lepiovre basal medium supplemented with 16 g l⁻¹ glucose, 0.5 g l⁻¹ AgNO₃, 2.2 mg l⁻¹ thidiazuron (TDZ), 0.5 mg l⁻¹ α-naphthaleneacetic acid (NAA), 2.5 g l⁻¹ Gelrite, seeds germinated in dark for 3 days and explants cultured in light, gave the best regeneration frequency (over 95%). The results also suggest that reducing the content of NH₄⁺ or keeping a suitable NO₃⁻/NH₄⁺ ratio in the regeneration medium would be crucial to Crambe shoot regeneration.     

Antifungal activities of extracts produced by liquid fermentations of Chilean Stereum species against Botrytis cinerea (grey mould agent)

Extracts obtained from liquid mycelial fermentations of Chilean species of the Stereum genus showed antifungal activity against Botrytis cinerea. Thirty-six strains were evaluated in vivo and in vitro assays, 25 belonging to Stereum hirsutum (Sh) and 11 to Stereum rameale (Sr). Two types of extracts were obtained: EtOAc-extract (liquid phase) and MeOH-extract (mycelial phase). Plate diffusion assay showed that EtOAc-extracts were more active than MeOH-extracts. S. hirsutum included 4 strains with the highest antifungal activity (Sh134-11, Sh144-11, Sh152-11, Sh155-11) and S. rameale highlighted with two strains (Sr25-11, Sr27-11). Effects on the mycelial growth of B. cinerea showed that EtOAc-extracts produced by S. hirsutum (Sh134-11, Sh152-11) inhibited from 1000 μg mL⁻¹, reaching 67 and 49%, respectively. At 2000 μg mL⁻¹, these strains inhibited nearly 80% of mycelial growth. EtOAc-extract of Sh134-11 was more effective to control the sporogenesis, inhibiting in 100% the sporulation at 500 μg mL⁻¹. Assays showed that Sh134-11 and Sh152-11 exhibited a minimal fungicidal concentration (MFC) of 50 and 100 μg mL⁻¹ respectively and minimal inhibitory concentration (MIC) at 20 μg mL⁻¹. EtOAc-extracts of Sr25-11 and Sr27-11 showed MFC value at 100 μg mL⁻¹ and MIC at 20 and 50 μg mL⁻¹, respectively. Strawberries treated with 1000 μg mL⁻¹of Sh134-11 and Sr25-11 reached 82 and 72% of decay inhibition, respectively. Treatments with 2000 μg mL⁻¹ showed a decay inhibition of 90% approximately. In vivo tests are in accordance with the results obtained in vitro assays, confirming the efficacy of Sh134-11 and Sr25-11 to control B. cinerea. Differences in antifungal activities observed in the different strains suggested that the ability to produce bioactive compounds is not homogenously distributed among S. hirsutum and S. rameale. Our study would suggest that submerged fermentations of Chilean S. hirsutum strain Sh134-11 produce extracts, which could be used as possible biofungicides and an alternative to synthetic fungicides.     

Optimization of ultrasonic-assisted extraction for herbicidal activity of chicory root extracts

The present study evaluated the herbicidal potential of extracts from chicory (Cichorium intybus L.) roots on the germination of Echinochloa crusgalli L. Beauv and Amaranthus retroflexus L. Eight ultrasonic assisted-extraction (UAE) conditions were optimized, using an orthogonal matrix design. The extract concentrations that would yield the largest allelopathic effects on the plant species were estimated by a modeling analysis. Our results showed that an alcohol solvent extract of chicory root had significant herbicidal activity which depended on the extract concentrations and the target species. The half-inhibitory concentrations of crude extract of chicory root ranged from 0.5 g l⁻¹ to 40.5 g l⁻¹. At a frequency of 40 kHz, the optimum UAE conditions to produce an extract for use as herbicide against E. crusgalli L. included an ethanol content (Ec) of 50% (v/v), a solvent-to-solid ratio (SR) of 16:1, an ultrasound temperature (UT) of 35 °C, an impregnation time (Imt) of 24 h with two rounds of impregnation (Imr), a sonication period (St) of 120 min with two rounds of sonication (Sr) and an ultrasound input power (P) of 200 W. The optimum conditions to produce an extract for use against A. retroflexus L. included an Ec of 100% (v/v), a SR of 16:1, an UT of 20 °C, an Imt of 48 h with two Imr, a St of 30 min with one Sr and a P of 400 W. The extract had the largest inhibitory effects on the germination index and root growth of both E. crusgalli L. and A. retroflexus L. at concentrations ranging from 30.8 to 33.7 g l⁻¹. At a concentration of 4.2 g l⁻¹, the extract significantly enhanced the shoot growth of A. retroflexus L. Overall, chicory root extract has potential for use as a main ingredient in natural herbicides or for development as a novel plant-derived herbicide.     

Improvement of growth and gymnemic acid production by altering the macro elements concentration and nitrogen source supply in cell suspension cultures of Gymnema sylvestre R. Br

Gymnema sylvestre is an important medicinal plant which bears bioactive compound namely gymnemic acids. The present work deals with optimization of cell suspension culture system of G. sylvestre for the production of biomass and gymnemic acid and we investigated effects of macro elements (NH₄NO₃, KNO₃, CaCl₂, MgSO₄ and KH₂PO₄ - 0.0, 0.5, 1.0, 1.5 and 2.0× strength) and nitrogen source [NH₄⁺/NO₃⁻ ratio of: 0.00/18.80, 7.19/18.80, 14.38/18.80, 21.57/18.80, 28.75/18.80, 14.38/0.00, 14.38/9.40, 14.38/18.80, 14.38/28.20 and 14.38/37.60 (mM)] of Murashige and Skoog medium on accumulation of biomass and gymnemic acid content. The highest accumulation of biomass (165.00 g l⁻¹ FW and 15.42 g l⁻¹ DW) was recorded in the medium with 0.5× concentration of NH₄NO₃ and the highest production of gymnemic acid content was recorded in the medium with 2.0× KH₂PO₄ (11.32 mg g⁻¹ DW). The NH₄⁺/NO₃⁻ ratio also influenced cell growth and gymnemic acid production; both parameters were greater when the NO₃⁻ concentration was higher than that of NH₄⁺. Maximum biomass growth (159.72 g l⁻¹ of FW and 14.95 g l⁻¹ of DW) was achieved at an NH₄⁺/NO₃⁻ ratio of 7.19/18.80, and gymnemic acid production was also greatest at the same concentration of NH₄⁺/NO₃⁻ ratio (11.35 mg g⁻¹ DW).     

Actinomycetes mediated biochemical responses in tomato (Solanum lycopersicum) enhances bioprotection against Rhizoctonia solani

Six actinomycetes isolates, namely Streptomyces toxytricini vh6, Streptomyces flavotricini vh8, S. toxytricini vh22, Streptomyces avidinii vh32, Streptomyces tricolor vh85 and vh41, an isolate of an unknown species of Actinomycetales, were tested for their efficacy in protecting tomato (Solanum lycopersicum) against Rhizoctonia solani under green house conditions. Actinomycetes treated plants showed better growth in terms of high chlorophyll content, higher phenylalanine ammonia lyase (PAL) activity and high total phenolic content. Qualitative and quantitative estimation of phenolic compounds from tomato leaves showed significant accumulation of six phenolic acids, gallic (29.02 μg g⁻¹ fresh leaf wt), ferulic (11.44 μg g⁻¹ fresh wt), cinnamic (56.84 μg g⁻¹ fresh wt), gentisic (24.19 μg g⁻¹ fresh wt), chlorogenic acid (1.72 μg g⁻¹ fresh wt) and salicylic (0.39 μg g⁻¹ fresh wt) acid, in actinomycetes treated plants. Biochemical profiling, when correlated with plant mortality in actinomycetes treated and untreated plants, indicated that isolates vh6 and vh8 offered 44.55% and 40.14% disease reductions, respectively compared to the control. These results established that these organisms have the potential to act as biocontrol agents.     

In vitro antifungal activity of extracts of Mexican Chihuahuan Desert plants against postharvest fruit fungi

Chemical fungicides have been intensively used in the control of diseases in fruits in postharvest conditions; nevertheless these actions have developed resistance in the phytopathogens, have contaminated the atmosphere, and have affected people's health through residual toxic compounds present in the food for human consumption. The objective of this study was to evaluate the in vitro inhibiting activity of extracts of Lippia graveolens Kunth, Agave lechuguilla Torr. Yucca carnerosana (Trel.) Mc Kelvey and Yucca filifera Chaub., against Rhizopus stolonifer, Colletotricum gloesporoides and Penicillium digitatum the postharvest fungi that causes diseases in fruits of commercial interest. The plants were collected in the neighborhood of the cities of Saltillo and Torreon in the Southern Region of the State of Coahuila in the semi-arid zones of the North of Mexico. The extracts of the plants were obtained with ethanol and hexane as solvents, and the resin was separated of solvent in a rotary evaporator. The fungi were isolated of apple, mango and orange fruits and were then seeded in Potato-Dextrose-Agar medium; the bioassays were inoculated with fungi and developed in Petri dishes. The evaluated variables were: Inhibition of mycelial growth, and Inhibition of the number of spores. For the analysis of the information a completely randomized experimental design in factorial arrangement was used. The results showed that the ethanolic extracts of L. graveolens at 500 μl L⁻¹ presented a fungicide effect of 100% against R. stolonifer, Y. filifera at 4000 μl L⁻¹ inhibited 100% the pathogen. The hexanic extracts of L. graveolens presented an inhibition of 100% at 500 μl L⁻¹. A. lechuguilla inhibited the pathogen in 100% at 3000 μl L⁻¹. The extract of L. graveolens, A. lechugilla, Y. carnerosana and Y. filifera showed its inhibiting activity in different percentages against the development of R. stolonifer, Colletotrichum gloeosporioides, and P. digitatum. This is the first scientific report that shows anti-fungal activity of ethanolic and hexanic extracts of L. graveolens, A. lechuguilla, Y. carnerosana, and Y. filifera against R. stolonifer, C. gloesporoides, and P. digitatum.     

Fungicidal activity of fluopicolide for suppression of Phytophthora capsici on squash

The efficacy of a new fungicide fluopicolide in suppression of Phytophthora blight caused by Phytophthora capsici was evaluated under laboratory and field conditions. Studies with 51 P. capsici isolates from vegetable crops in Georgia, USA, indicated that 5.9% of the isolates were resistant, 19.6% were intermediately sensitive, and 74.5% were sensitive to 100 μg ml⁻¹ of mefenoxam based on in vitro mycelial growth. EC₅₀ values of fluopicolide in inhibiting mycelial growth of 25 isolates, representing resistant, intermediately sensitive, and sensitive to mefenoxam, ranged from 0.05 to 0.35 μg ml⁻¹ with an average of 0.2 μg ml⁻¹ EC₅₀ values of fluopicolide in suppressing zoospore germination and sporangium production of the 25 isolates ranged from 1.1 to 4.5 μg ml⁻¹ and 0.3–9.0 μg ml⁻¹, respectively. Evaluation of a collection of 150 P. capsici isolates from vegetables and irrigation ponds found none of the isolates were resistant to 10 μg ml⁻¹ of fluopicolide. Field experiments were conducted to determine the efficacy and application methods of fluopicolide for control of P. capsici on squash in spring 2007 and 2009. Fluopicolide applied through drip irrigation or as a foliar spray at 86.6 or 115.4 g ha⁻¹ consistently provided significant disease reduction and increased squash yield. Results with fluopicolide were similar or slightly superior to mefenoxam applied at recommended rate. Fluopicolide applied at 57.7 g ha⁻¹ did not provide consistent satisfactory disease suppression. The results indicated that fluopicolide was effective in suppression of different stages of the life cycle of P. capsici and could be a viable alternative to mefenoxam for managing Phytophthora blight in squash production.     

Amaranth (Amaranthus hypochondriacus) as an alternative crop for sustainable food production: Phenolic acids and flavonoids with potential impact on its nutraceutical quality

The demand for food is increasing, not only to meet food security for growing populations, but also to provide more nutritious food, rich in good quality proteins and nutraceutical compounds. The amaranth (Amaranthus hypochondriacus) plant, in addition to its high nutritive and nutraceutical characteristics, has excellent agronomic features. The objective of the present study was to analyze some physical and proximal-nutritional properties of amaranth seeds obtained from different varieties grown in arid zones and characterize their phenolic acids and flavonoids. Two commercial (Tulyehualco and Nutrisol) and two new (DGETA and Gabriela) varieties of A. hypochondriacus were grown at the Mexican Highlands zone. Tulyehualco and DGETA varieties had higher seed yield of 1475 and 1422 kg ha⁻¹, respectively, comparable to corn and soybean production in agricultural areas. Gabriela had the highest protein content of 17.3%, but all varieties had an adequate balance of essential amino acids. Polyphenols as rutin (4.0–10.2 μg g⁻¹ flour) and nicotiflorin (7.2–4.8 μg g⁻¹ flour) were detected. Amaranth can be cultivated in arid zones where commercial crops cannot be grown; the seeds besides their well known nutritive characteristics could be a source of phenolic compounds of high antioxidant properties.     

Chitosan enhances withanolides production in adventitious root cultures of Withania somnifera (L.) Dunal

Calli were obtained from leaf, cotyledon and internode explants of in vitro-grown plants of Indian cultivar of Withania somnifera in MS medium supplemented with 2, 4-D (2.0 mg l⁻¹) and Kinetin (0.2 mg l⁻¹). The brown, semi-friable callus (500 mg FW) derived from leaf explants produced higher number of primary adventitious roots (9 roots/callus) in half strength MS medium fortified with IBA (0.5 mg l⁻¹) and NAA (0.1 mg l⁻¹). The primary adventitious roots with an inoculum mass of 15 g FW were cultured for 6 weeks in the same medium for secondary adventitious root proliferation. Elicitation of abiotic elicitor, aluminium chloride at 10 mg l⁻¹ at the end of 4 weeks culture with 4 h exposure time enhanced withanolides productivity. Under similar culture conditions, the biotic elicitor, chitosan at 100 mg l⁻¹ stimulated higher production of all withanolides when compared to aluminium chloride treatment. This is the first report on the use of callus-derived adventitious root culture for the enhanced production of withanolides upon chitosan elicitation.     

Insecticidal activities of crude extracts and phospholipids from Chenopodium ficifolium against melon and cotton aphid, Aphis gossypii

Several organic solvent extracts of Chenopodium ficifolium were tested for their insecticidal activity against melon and cotton aphid, Aphis gossypii, on cucumber plants. Both methanol and ethanol extracts, at 5000 μg ml⁻¹, were highly active giving over 80% control. The other crude extracts displayed moderate or weak insecticidal activity giving control in the range of 16–69%. Two phospholipids were isolated as insecticidal active substances from C. ficifolium. Their chemical structures were identified as 1-palmitoyl-2-(3-trans)-hexadecenoyl-sn-glycero-3-phosphoglycerol and 1-palmitoyl-2-linoleoyl-3-glycerophosphocholine by GC–MS, EDS, mass and NMR spectral analyses. Both compounds displayed a dose-dependent mortality of A. gossypii. Furthermore, the liquid formulation that was obtained by partitioning with n-hexane from the methanol extract of C. ficifolium controlled melon and cotton aphid on cucumber plants effectively. These results indicate that extracts of C. ficifolium have potential for development as botanical insecticides for controlling A. gossypii infesting cucumber plants.