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The stem bark of Geoffroea decorticans(Gill.ex Hook. etArn.) Burk. was used medicinally to cure several skin affections; however, phytochemical and biological antecedents were not found. Analyses of purified methanolic extract from G. decorticansbark (PFGB), realized by silylation derivatization for GC/MS, C18-CC and HPLC followed by two-dimensional TLC and UV-Vis spectroscopy, allowed to characterize nine phenolic compounds, among these, two methoxy flavonoids. Antibacterial assays of PFGB showed the highest activity (MICs= 125 μg/mL) against Staphylococcus aureus(25923) and Enterococcus faecalis(29212) ATCC strains. Moreover, PFGB showed the highest intracellular antioxidant activity at low concentration (5 μg/mL), evaluated by using the fluorescent DA-H2DCF probe on lymphocyte culture; cytotoxic effects on lymphocytes activated or not by LPS were not observed, through Trypan Blue Exclusion and MTT colorimetric assays.The results obtained from the ethnomedicinal approach of this work contribute to the scientific validation of the vulnerary medicinal use of G. decorticans.

A fraction enriched in triterpenoid saponins (F4) from Anagallis arvensis L. was chemically characterized by UHPLC-ESI-QTOF-MS/MS and NMR analyses. The results proposed the presence of nine monodesmosidic saponins derived from oleanolic acid, including two reported for the first time for this species, 3: 3-O-{β-D-glucopyranosyl (1→4)-[β-D-xylopyranosyl (1→2)]-β-D-glucopyranosyl (1→4)-[β-D-glucopyranosyl (1→2)]-α-L-arabinopyranosyl}-13β, 28-epoxy- 22 acetyl- 28 methoxy-16α, 24-oleananediol and 4: 3-O-{β-D-glucopyranosyl (1→4)-β-D-glucopyranosyl (1→4)-[β-D-xylopyranosyl (1→2)]-β-D-glucopyranosyl (1→4)-[β-D-glucopyranosyl (1→2)]-α-L-arabinopyranosyl}-13β, 28-epoxy- 16α, 30-oleananediol. Furthermore, haemolytic activity was determined by dot-blot autography, and cytotoxicity on human lymphocyte cultures was analysed according to metabolic activity (MTT assay) and membrane integrity (Trypan blue exclusion test). F4 showed mild cytotoxicity (%V > 50% at 100 μg/mL) on human lymphocytes under conditions of activation or not by LPS; moreover, showed haemolytic activity between 50 and 500 μg. This work contributed to phytochemical knowledge of the triterpenoid saponins from A. arvensis, and its cytotoxic effects on normal human cells.

The antiochratoxigenic (anti-OTA) activity of dichloromethane extract of Macfadyena cynanchoides stems, its antifungal metabolites (lapachol and 1-hydroxy-4-methyl anthraquinone) and sodium metabisulfite was evaluated alone and in combination with this commercial preservative on Aspergillus species at sub-lethal concentrations. Grape juice inoculated with 5×103 spores/mL of A. carbonarius or A. niger was supplemented with the extract (2500 to 156.3 µg/mL), the quinones (1250 to 78.1 µg/mL) or sodium metabisulfite (2500 to 156.3 µg/mL). OTA accumulation was measured in the sub-lethal concentrations after incubating in the dark for 6 days at 15°C. The extract and sodium metabisulfite completely inhibited growth of both fungi at 2500 µg/mL while lapachol and 1H4MA required 1250 µg/mL. Average OTA biosynthesis declined 30-60% respect to control at extract concentrations of 625-1250 µg/mL. Both lapachol and 1H4MA tested at 625 µg/mL exerted a similar anti-OTA effect with an average inhibition of 30%. In the case of sodium metabisulfite, it increased OTA production between 60 to 100% at all the sub-lethal concentrations. Binary mixtures of extract or lapachol or 1H4MA + sodium metabisulfite tested at concentrations of 9.8 + 156.3 µg/mL, 19.5 + 312.5 µg/mL and 39.1 + 625 µg/mL completely inhibited OTA production. In addition, the mixture lapachol + sodium metabisulfite (9.8 + 156.3 µg/mL) delayed a little the time required for completion of the alcoholic fermentation, but did not reduce the Saccharomyces cerevisiae viability or modify physico-chemical parameters measured at the end of the process. Our results show that the mixture sodium metabisulfite + lapachol has potential as an antifungal and anti-OTA agent useful to reduce the doses of sulfites in the control of Aspergillus on post-harvested grapes.

Essential oils from aerial parts of the herbs Peganum harmala and Nepeta cataria, and leaves of the tree Phellodendron amurense were analyzed by GC-FID and GC-MS, and their larvicidal activities were assayed on the early fourth instar larvae of Aedes aegypti. The major constituents of the oils were limonene (14.5%) and thymol (11.5%) in P. harmala, thymol (46.5%), 4aα,7α,7aβ-nepetalactone (18.3%) and 4aα,7β,7aα-neptalactone (19.7%) in N. cataria, eugenol (14.5%) andγ-eudesmol (9.5%) in P. amurense.The oil of N. cataria had a strong larvicidal activity (LC50 < 50 µg/mL; LC90 < 86.8 µg/mL) on A. aegypti while the remaining oils showed a moderated killing effect. The larvicidal activity of N. cataria oil was associated to the contents of 1,8-cineol, camphor, 4aα,7α,7aβ-Nepetalactone, 4aα,7β,7aα-Nepetalactone and thymol. Our results indicate that the oil of N. catariadeserves to be used as a source of larvicidal agents against A. aegypti. Fil: Shengxiang, Yang. Zhejiang A & F University; China Fil: Mingsheng, Bai. Ningxia University; China Fil: Jian Yiang. China Academy Of Chinese Medical Sciences; China Fil: Yuang, Yuang. China Academy Of Chinese Medical Sciences; China Fil: Yamei, Zhang. Jilin University; China Fil: Jianchun, Qin. Jilin University; China Fil: Yi, Kuang. Zhejiang A & F University; China Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina

The essential oil from the aerial parts of Lobularia maritima was investigated for its chemical composition, and its repellent and insecticidal efficacy against the grain pests Callosobruchus maculatus, Tribolium castaneum and Sitophilus oryzae. A number of 41 compounds were identified by GC-MS from which azeleonitrile (39.7%), trans-3-pentenenitrile (36.3%) and 4-isothiocyanato1-butene (10.9%) were the most abundant. A fumigant bioassay-guided fractionation of the essential oil constituents led to the isolation of trans-3-pentenenitrile. Its structure was confirmed by EI-MS and NMR techniques. Fumigant effect of the essential oil was strong (LC50 = 7.48 μL/L) on C. maculatum and moderate on S. oryzae and T. castaneum (LC50 = 35.37 and 59.94 μL/L, respectively). trans-3-pentenenitrile showed strong fumigant effect on the three pest species (LC50 = 6.62–8.36 μL/L). Both the oil and trans-3-pentenenitrile showed strong contact effect with LD50 values in the range 4.84–7.81 µg/adult. The oil showed a repellency of 100% on C. maculatus and S. oryzae at concentrations higher than 0.05 and 0.1 nL/cm2 , respectively, and 93% against T. castaneum yet at 0.2 nL/cm2 . trans-3-pentenenitrile also showed 100% repellency against C. maculatus (≥ 0.05 nL/cm2 ), S. oryzae (≥ 0.15 nL/cm2 ) and T. castaneum (0.2 nL/cm2 ). The results in this study indicated that the essential oil of L. maritima is an important source of trans-3-pentenenitrile which can be used in the development of insecticidal agents against the three grain pests. Fil: Wang, H. Y.. Jilin University; China Fil: Zhang, M. Z.. Jilin University; China Fil: Han, X.. Jilin University; China Fil: Cong, J.. Jilin University; China Fil: Wang, S. Q.. Jilin University; China Fil: He, S. L.. Jilin University; China Fil: Wei, D. S.. Jilin University; China Fil: Zhang, Y. B.. Jilin University; China Fil: Qin, Jianchun. Jilin University; China Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina

Fungal co-culture has several biotechnological applications including the discovery or the enhanced production of secondary metabolites. It is also a powerful tool aiding to elucidate the involvement of secondary metabolism in fungus-fungus interactions. The aim of this work was to investigate secondary metabolites produced when Fusarium verticillioides is co-cultured with Gliocladium roseum. Secreted metabolites were analyzed by HPLC-MS, and fusaric acid (FA) was quantified by HPLC-DAD. Four FA derivatives were identified only in the F. verticillioides-G. roseum co-culture. Mass spectrometry and one- and two-dimensional NMR spectra indicated that they were 5-butylpyridine-2-carboxylic acid methyl ester (5B2CAM), 4-(5-butylpicolinamido) butanoic acid (45BBA), methyl 4-(5-butylpicolinamido) butanoate (M45BBA), and bis(5-butyl-2-pyridinecarboxylate-N1,O2)-copper (B52P). 45BBA and M45BBA are reported for the first time and were FA biotransformation products generated by G. roseum. The antifungal activity of 5B2CAM, 45BBA, and M45BBA was evaluated in vitro against Botrytis cinerea and Aspergillus niger. They were less fungitoxic than FA, with 45BBA as the least toxic. Our results suggest that the effective antagonism exerted by G. roseum against F. verticillioides is due, at least in part, to its detoxifying ability against FA.

Mapping QTL for disease resistance and associated traits is important to develop maize (Zea mays L.) hybrids less susceptible to ear rots. A biparental mapping population of 298 F5 recombinant inbreds (RIs), obtained from the cross between LP4637 (moderately resistant) and L4674 (susceptible), was genotyped with 250 single nucleotide polymorphism (SNP) markers. A set of 120 of those RIs, selected by uniRec procedure, and parental inbreds were phenotyped in two environments for pericarp thickness, pericarp content of trans-ferulic acid (tFA) and resistance to Fusarium ear rot. The set of parental inbreds exhibited an average density of one marker every 5 cM, 6% of a residual heterozygosity, and 5% of lost data. Moderate negative genotypic correlations were observed between disease severity and pericarp thickness (rG = − 0.31) and between disease severity and pericarp content of tFA (rG = − 0.32). Quantitative trait loci were mapped for disease severity in bins 1.06, 2.03, 3.06, 5.04, 5.07 and 6.05, for pericarp thickness in bins 1.06, 2.03, 4.02 and 4.05, and for pericarp content of tFA in bins 2.03, 3.06, 4.05 and 6.05. The joint models, including some additive-by-additive epistasis gene effects, explained 56.0–58.2%, 46.6–45.5%, 41.4–47.1% of the phenotypic variability for disease severity, pericarp thickness and pericarp content of tFA, respectively, depending on the environment. The most important QTL for the three traits overlapped in bin 2.03 indicating that genes from this genomic region might contribute to the expression of disease resistance by increasing thickness and tFA content of the pericarp.

Aspidosperma quebracho-blanco Schltdl. is a tree that - at maturity - can reach up to 25 m in height. It has a straight trunk that can reach 0.8–1 m in diameter. It is commonly known as “quebracho blanco” and widely used in the Chaco region as medicine to treat fever, malaria, swellings, stomach upsets, cough, headaches, syphilis, impotence, benign prostatic hypertrophy and asthma related dyspnea. The stem bark of A. quebracho-blanco is the most frequently used plant part in medicinal preparations. Terpenoid indole alkaloids, the main biologically active principles of the plant are associated with the medicinal properties of the tree. The bark is rich in aspidospermine, yohimbine, deacetyl-aspidospermine and quebrachamine. Other alkaloids also were isolated. Several of these terpenoid indole alkaloids showed in vitro and sometimes in vivo pharmacological activities that support the medicinal effects traditionally attributed to A. quebracho-blanco. Aspidospermine showed in vitro antiplasmodial activity. Aspidospermine, yohimbine and quebrachime had adrenergic blocking activities on a variety of urogenital tissues. Rhazinilam showed mild analgesic activity in mice. The high tannin levels found in the bark, leaves and woods are believed to contribute to the antioxidant and anti-inflammatory activities. Recent research suggests that bark preparations have a potential for the development of antiviral and larvicidal agents. However, further investigations are needed with especial emphasis on the in vivo effects, in order to understand the real therapeutic significance of the alkaloids and other compounds of A. quebracho-blanco.

Lippia integrifolia (Griseb.) Hieron. is a native aromatic shrub, commonly known as “incayuyo” or “inca tea”. It is widely used in traditional medicine in northern and central Argentina, where infusions of leaves and flowers are used as digestive, emmenagogue, antitussive, carminative, and against dyspepsia, stomachache, flu and indigestions. Its essential oil is a rich source of sesquiterpenoids based on unique or very rare skeletons such as integrifoliane, lippifoliane, asteriscane, africanane and 4,5-seco-africanane. The essential oil shows a remarkable chemical diversity, with at least five clearly defined chemotypes. Two of the most frequently found chemotypes stand out for their unique components, one called “trans-davanone chemotype” that has this ketone as the dominant component (>75%); and the other, called “lippifolienone chemotype”, rich in oxygenated sesquiterpenes based on the rare lippifoliane and integrifoliane skeletons. Aqueous and ethanol extracts of “incayuyo” showed antiadhesive properties against Helicobacter pylori. In addition, the secretion of IL-8 induced by Helicobacter pylori was significantly reduced by co-incubation of AGS cells with “incayuyo” extract. Infusions and decoctions of “incayuyo” displayed strong antioxidant, choleretic and antispasmodic effects, stimulated phagocytosis rate of macrophages and inhibited the LPS-induced NO-secretion. In addition, a significant reduction of contractions induced by acetylcholine, CaCl2 and KCl was observed in isolated rat jejunum. The bioactive components of “incayuyo” extracts are flavonoid glycosides, (poly)-phenolic compounds, phenylethanoid glycosides (acteoside, isoacteoside) and sesquiterpenoids. All experimental data support the use of “incayuyo” in traditional medicine. Comparative studies on the pharmacological efficacy of the different essential oil chemotypes are necessary. The qualitative and quantitative content of non-volatile bioactive metabolites should be used to define quality parameters.

Xenophyllum poposum (Philippi) V.A. Funk (syn. Werneria poposa Philippi) is a creeping sub-shrub, 3–5 cm tall with an unpleasant smell, commonly known as “poposa” or “pupusa”. It is widely used in Andean traditional medicine to treat hypertension, altitude sickness (mountain sickness), hepatic diseases and digestive disorders. Mono- and sesquiterpenoids accompanied by benzofuran derivatives, such as tremetone and 6-hydroxytremetone, have been identified in its essential oil while p-hydroxyacetophenone and benzofuran derivatives, flavonoids, coumarins and diterpenoids, were isolated from organic extracts of aerial parts. 4-Hydroxy-3-(isopenten-2-yl)-acetophenone (1) is by far the dominant component of the methanol extract along with minor amounts of 6-hydroxytremetone (2); both compounds showing a hypotensive effect on rats and mice. Two components of its essential oil, namely, Τ-cadinol (10-epi-α-cadinol) and α-cadinol possess significant anti-inflammatory activity and inhibit nitric oxide production. All available information seems to support the uses of X. poposum in traditional medicine. Due to its numerous bioactive metabolites, strong effects should be expected after the consumption of this herb. Consequently, the dosage (amount consumed) should be carefully controlled to avoid undesirable effects. Further in-depth investigations are needed to chemically characterize the water soluble components of this plant and to identify possible chemotypes, as well as define quality parameters.

Lippia turbinata Griseb. is a native aromatic shrub commonly known as “poleo”, “te del pais”, “te criollo” or “poleo fino”. It is widely used in traditional medicine of Northern and central Argentina. Infusions of aerial parts are used as diuretic, tonic, emmenagogue, for dysmenorrhea, as a regulator of the menstrual cycle and as abortifacient. Its essential oil shows remarkable variability in chemical composition. Very little research has been conducted on its non-volatile components. Eight oleanane-type triterpenes were identified in a methanol-methylene chloride extract of “poleo”. There are also relatively few studies on the biological activity of this herb. Antifungal, antimicrobial, nematocidal and antiviral activities against the Junin-virus have been reported for the essential oil. Piperitenone oxide, one of the main components in several collections, has been evaluated in relation to the following biological activities: cardiovascular, hypotensive, bradycardic, insecticidal, trypanocidal, schistosomicidal, antimicrobial, antinociceptive, analgesic and larvicidal. Infusions are used to stimulate uterine motility and therefore should not be consumed by pregnant women. In order to better assess its pharmacological utility, much additional research is needed on the non-volatile metabolites and biological properties of this species.

Tessaria dodoneifolia [Asteraceae] is traditionally employed in Northwestern Argentina for fungal infections treatment. We report the antifungal activity guided isolation and identification of substances from aerial parts of this species, both individually and in combination with fluconazole (FLU), against Candida albicans strains. Two antifungal flavanones were identified as naringenin (NAR) and pinocembrin (PIN). These compounds could individually inhibit the growth of C. albicans strains. Combinations of NAR and PIN with FLU were synergistic against the FLU resistant and sensitive C. albicans strains. Genotoxic and cytotoxic evaluations were also performed. NAR, PIN and their combinations with FLU did not have a genotoxic effect on Bacillus subtilis rec strains. Finally, these compounds did not show cytotoxicity at concentrations below 80 μg/mL., Plant biology; Toxicology; Bioactive plant product; Mycology; Secondary metabolite; Flavonoid; Chromatography; Spectroscopy; Tessaria dodoneifolia; Antifungal activity; Candida albicans; Synergistic effect; Toxicity assays.

This work aimed to identify secondary metabolites from aerial parts of Euphorbia species functional for control of toxigenic Fusarium species responsible of cereal grain rots. Methods and Results: Aerial parts of Euphorbia serpens, Euphorbia schickendantzii and Euphorbia collina were sequentially extracted with hexane, ethyl acetate and methanol. The extracts were tested against strains of Fusarium verticillioides and Fusarium graminearum by microdilution tests. The hexane extract of E. collina provided the lowest IC50s on both fungal species. Further fractionation showed that cycloartenol (CA) and 24-methylenecycloartanol are associated to the moderate inhibitory effect of the hexane extract on fungal growth.Sublethal concentrations of CA and 24MCA blocked deoxynivalenol (DON) and fumonisins production.CA and 24MCA co-applied with potassium sorbate, a food preservative used for Fusarium control, synergized the growth inhibition of fungi. The mixtures reduced mycotoxins accumulation when applied at sublethal concentrations. Conclusions: CA and 24MCA inhibited both fungal growth and mycotoxins production. This fact is an advantage respect to potassium sorbate which increased the mycotoxins accumulation at sublethal concentrations. Significance and Impact of the Study: CA and 24MCA synergized potassium sorbate and their mixtures offer a lower mycotoxigenic risk than potassium sorbate for control of the Fusarium species. Fil: Jiménez, Cristina Marisol. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Alvarez, Hebe Lina. Universidad Nacional de La Pampa. Facultad de ciencias veterinarias; Argentina. Departamento de Epizootiologia y Salud Publica ; Facultad de Ciencias Veterinarias ; Universidad Nacional de la Pampa; Fil: Ballari, María Sol. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina Fil: Labadié, Guillermo R.. Universidad Nacional de Rosario; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Toso, Ricardo Enrique. Universidad Nacional de La Pampa; Argentina Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina

Erwinia carotovora subsp. cause the potato soft rot, which is a major disease in agriculture. Antibacterial agents currently applied on potato soft rot often offer a restricted control and have several disadvantages. Propolis has shown a wide range of antimicrobial activity, although its effect has not been investigated on E. carotovora subsp. In this work, we tested extracts from propolis samples of Northwest Argentina against E. carotovora subsp.RESULTS: Ethanolic propolis extracts (EPEs) from samples of Santiago del Estero province, particularly from sample 4 (EPE4), showed the highest antibacterial activity, which was associated with the highest content of flavonoids. 2′,4′‐Dihydroxychalcone, 2′,4′‐dihydroxy‐3′‐methoxychalcone, galangin, and pinocembrin were identified as antibacterial constituents of EPE4. 2′,4′‐Dihydroxychalcone showed an antibacterial activity (minimum inhibitory concentration, MIC = 0.3?1.2 μg gallic acid equivalents (GAE) mL−1; minimum bactericidal concentration, MBC = 0.6?4.8 μg GAE mL−1) lower than that of bacterimycin (MIC = 2.4?9.6 μg mL−1; MBC = 19.2?38.4 μg GAE mL−1) and streptocycline (MIC = 19.2?38.4 μg mL−1; MBC = 38.4?76.8 μg mL−1). Preventive assays on unwounded and wounded potatoes showed that their immersion in EPE4 containing 87.5 μg GAE mL−1 or streptocycline containing 40 μg mL−1 was equally effective in controlling potato soft rot, reducing the disease incidence by 64.6?67.0% (unwounded tubers) and 88.0?86.0% (wounded tubers) and the disease severity by 49.8?49.8% (unwounded tubers) and 54.5?68.5% (wounded tubers).CONCLUSIONS: Flavonoid‐rich propolis extracts from Northwest Argentina efficiently reduced in vivo the incidence and severity of potato soft rot caused by E. carotovora subsp. Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Bertini Sampietro, Maria Sofia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina

The trees Schinopsis lorentzii and S. marginata produce alk(en)ylcatechols, also known as urushiols, which function in plant defense as antifungals, antifeedants and cause contact dermatitis in human beings. Here, we propose the use of the diazonium salts Fast Blue B ½ZnCl2 and Fast Blue RR ZnCl2 for the in situ histochemical localization of urushiols in stems and leaflets of S. lorentzii and S. marginata. The main surface and internal anatomical features of the aerial plant parts were visualized with cresyl violet and astra blue safranin. Extracts from leaflets and stems were separated by TLC and revealed with aqueous solutions of 0.5% Fast Blue B ½ZnCl2 in 5% acetic acid (FBB), 0.5% Fast Blue RR ZnCl2 in 5% KOH (FBRR) and 5% FeCl3. Fresh cross sections of leaflets and stems were also stained with these chromogenic reagents. Mature leaflets and stems showed few glandular trichomes in their surfaces. Well developed phloem schizogenous ducts were uniformly distributed in the leaflet mesophylls and radially organized in the cross sections of the stems. TLC of the extracts revealed with FBB, FBRR and 5% FeCl3 gave bluish black spots for urushiols. Very few constituents of the extracts other than the urushiols revealed the same colour after spraying with the diazonium salts. FBB and FBRR indicated the presence of urushiols in both the cavity of the phloem schizogenous ducts and their surrounding epithelial cells. Urushiols were not visualized neither in the cuticle of the aerial parts nor in the glandular trichomes. FBB was coupled to more individual polyphenol species than FBRR whereas FeCl3 was by far a less specific chromogenic reagent. Nevertheless, FBRR was more selective towards the urushiols than FBB. The exposure to urushiols should be possible only after the breakdown of leaflets and stems of Schinopsis trees. Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Mercado, Maria Ines. Fundación Miguel Lillo; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico - Tucumán. Unidad Ejecutora Lillo; Argentina Fil: Aristimuño Ficoseco, Maria Eugenia del Milagro. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Ponessa, Graciela. Fundación Miguel Lillo; Argentina Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina

Ethnopharmacological relevance Anagallis arvensis L. (Primulaceae) is used in argentinean northwestern traditional medicine to treat fungal infections. We are reporting the isolation and identification of compounds with antifungal activity against human pathogenic yeast Candida albicans, and toxicity evaluation. Aim of the study to study the antifungal activity of extracts and purified compounds obtained form A. arvensis aerial parts, alone and in combinations with fluconazole (FLU), and to study the toxicity of the active compounds. Materials and methods Disk diffusion assays were used to perform an activity-guided isolation of antifungal compounds from the aerial parts of A. arvensis. Broth dilution checkerboard and viable cell count assays were employed to determine the effects of samples and combinations of FLU + samples against Candida albicans. The chemical structures of active compounds were elucidated by spectroscopic analysis. Genotoxic and haemolytic effects of the isolated compounds were determined. Results Four triterpenoid saponins (1–4) were identified. Anagallisin C (AnC), exerted the highest inhibitory activity among the assayed compounds against C. albicans reference strain (ATCC 10231), with MIC-0 =1 µg/mL. The Fractional Inhibitory Concentration Index (FICI=0.129) indicated a synergistic effect between AnC (0.125 µg/mL) and FLU (0.031 µg/mL) against C. albicans ATCC 10231. AnC inhibited C. albicans 12–99 FLU resistant strain (MIC-0 =1 µg/mL), and the FICI=0.188 indicated a synergistic effect between AnC (0.125 µg/mL) and fluconazole (16 µg/mL). The combination AnC+ FLU exerted fungicidal activity against both C. albicans strains. AnC exerted inhibitory activity against C. albicans ATCC 10231 sessile cells (MIC50=0.5 µg/mL and MIC80=1 µg/mL) and against C. albicans 12–99 sessile cells (MIC50=0.75 µg/mL and MIC80=1.25 µg/mL). AnC exerted haemolytic effect against human red blood cells at 15 µg/mL and did not exerted genotoxic effect on Bacillus subtilis rec strains. Conclusions The antifungal activity and lack of genotoxic effects of AnC give support to the traditional use of A. arvensis as antifungal and makes AnC a compound of interest to expand the available antifungal drugs. Fil: Soberon, Jose Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Biológica. Cátedra de Fitoquímica; Argentina Fil: Sgariglia, Melina Araceli. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Biológica. Cátedra de Fitoquímica; Argentina Fil: Pastoriza, Ana Cristina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Biológica. Cátedra de Fitoquímica; Argentina Fil: Soruco, Estela M.. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Biológica. Cátedra de Fitoquímica; Argentina Fil: Jäger, Sebastián N.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina Fil: Labadie, Guillermo Roberto. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Química Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Química Rosario; Argentina. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas; Argentina Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucuman. Facultad de Bioquímica, Química y Farmacia. Instituto de Química Biológica. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

The leaf extracts of bamboo (Shibataea chinensis Nakai) and its fractions (ethyl acetate, n-butanol and water) obtained after extraction with petroleum ether were assayed on seed germination and seedling growth of lettuce and cucumber. The leaf extract showed the lowest phytotoxic activity while the n-butanol fraction was the most bioactive on both target plants. A bioassay guided isolation of the n-butanol fraction led to the isolation of five phytotoxic compounds. The chemical structures of these compounds were elucidated by nuclear magnetic resonance as apigenin (I), quercetin (II), daucosterol (III), 3,3´,4´,5-tetrahydroxy-7-methoxyflavone (IV) and p-methoxybenzoic acid (V). The quercetin, apigenin and daucosterol were more phytotoxic to germination (IC50=7.26-7.95 ppm), root (IC50=2.97-6.33 ppm) and shoot elongation (IC50=4.89-7.74 ppm) of lettuce than glyphosate (IC50>20 ppm). In cucumber, the p-methoxybenzoic acid showed similar phytotoxic activity (IC50=20 ppm, 6.93 ppm, 10.45 ppm, respectively) to glyphosate (IC50=20 ppm, 7.71 ppm, 10.84 pppm, respectively). Our research suggested that the leaves of S. chinensis have potent allelochemicals and their potential as herbicide should be further investigated. Fil: Kuang, Y.. Zhejiang A＆F University; China Fil: Yang, S. X.. Zhejiang A＆F University; China Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Zhang, X. F.. Zhejiang A＆F University; China Fil: Tan, J.. Zhejiang A＆F University; China Fil: Gao, Q. X.. Zhejiang A＆F University; China Fil: Liu, H. W.. Zhejiang A＆F University; China Fil: Ni, Q. X.. Zhejiang A＆F University; China Fil: Zhang, Y. Z.. Zhejiang A＆F University; China

Septoria brown spot and Cercospora leaf blight are late season diseases caused by Septoria glycines and Cercospora kikuchii, respectively. New antifungals are required against these diseases because the chemical controls currently used have detrimental impacts on wildlife and human health. In this work, 48 extracts originated from the leaves, bark, sapwood or heartwood of four forestry species were assayed by the disc diffusion method against S. glycines and C. kikuchii. Although 18 extracts showed antifungal activity, only 5 were active on both fungal species. The leaf methanolic extract of Blepharocalyx salicifolius showed the lowest minimum inhibitory dose (MID) and the highest diameter of growth inhibition (DI) on both fungal species (MID = 200 μg, DI = 14.2 mm, C. kikuchii; MID = 400 μg, DI = 12.2 mm, S. glycines). Pinocembrin was identified as the main antifungal constituent of the methanolic extract. Both the methanolic leaf extract of B. salicifolius and pinocembrin synergized in vitro the effect of the fungicide difenoconazole. Preventive applications of the extract and the mixture extract + difenoconazole (2.4 mg/mL + 0.006 mg/mL) strongly reduced disease severity generated by S. glycines and C. kikuchii 21 days after inoculation of the soybean plants. This effect was significantly stronger than that generated by difenoconazole. Our results suggest that the application of the methanolic extract of B. salicifolius, alone or in mixture with difenoconazole is a promising strategy to be incorporated in the chemical control of S. glycines and C. kikuchii. Fil: Sequin, Christian Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Entre Ríos; Argentina Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Gómez, Analia de Los Angeles. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Aceñolaza, Pablo Gilberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Entre Ríos; Argentina

Extracts from aerial parts of Prosopis ruscifolia, Bidens pilosa, Cercidium praecox and Phoradendron liga were assayed against toxigenic Aspergillus species. They were obtained by sequential extraction of the aerial parts with hexane (fHex), dichloromethane (fDCM), ethyl acetate (fEtOAc) and methanol (fMeOH). The fMeOH from P. ruscifolia showed the highest antifungal spectrum (MIC = 750–1500 µg mL−1; MID = 50–200 µg; DI = 1.7–3.0 mm). Indolizidine alkaloids (juliflorine and juliprosine) and tryptamine were identified with strong (MIC = 188 µg mL−1) and moderate antifungal activities (MIC = 750 µg mL−1), respectively, towards A. parasiticus and A. flavus. The fMeOH, the indolizidine alkaloids and tryptamine synergized the fungitoxic effect of potassium sorbate and propiconazole. They completely suppressed the biosynthesis of aflatoxins at concentrations of 47, 94 and 375 µg mL−1, respectively. Our results indicate that fMeOH and its identified alkaloids are promisory additives of commercial antifungals and are antiaflatoxigenic agents at concentrations below of those required for complete suppression of fungal growth. Fil: Gómez, Analia de Los Angeles. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Terán Baptista, Zareath Pamela. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Mandova ,Tzvetelina. 2laboratoire de Pharmacognosie, Umr/cnrs 8638 Comete, U; Francia Fil: Barouti,Angeliki. 2laboratoire de Pharmacognosie, Umr/cnrs 8638 Comete, U; Francia Fil: Kritsanida, Marina. 2laboratoire de Pharmacognosie, Umr/cnrs 8638 Comete, U; Francia Fil: Grougnet Raphael. 2laboratoire de Pharmacognosie, Umr/cnrs 8638 Comete, U; Francia Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales. Cátedra de Fitoquímica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina

Twenty four extracts from Bignoniaceae plants of northwest Argentina were tested for antifungal activity against

Twenty four extracts from Bignoniaceae plants of northwest Argentina were tested for antifungal activity against Aspergillus species responsible of the grape black rot. Stems and leaves of Amphilophium cynanchoides, Macfadyena cynanchoides, Tecoma stans and Jacaranda mimosifolia were separately extracted with solvents of increasing polarity to obtain the dichloromethane (fCH2Cl2), ethyl acetate (fEtOAc) and methanol extracts (fMeOH). The fCH2Cl2 from stem of M. cynanchoides had the lowest IC50 (1.0-1.2 mg/mL) and MID values (0.6-1.2 mg) and the highest ID values (5.0-6.8 mm) on A. niger and A. carbonarius. The main contributors of the antifungal activity of fCH2Cl2 were identified as lapachol (MIC = 0.25-1.00 mg/ml) and 1-hydroxy-4-methylanthraquinone (MIC = 0.0625-0.125 mg/mL). These compounds synergized the antifungal activity of sodium metabisulfite and showed an additive effect in mixtures with propiconazol. They might be used as additives of commercial antifungals to protect grapes against A. niger and A. carbonarius. Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Universidad San Pablo Tucumán; Argentina Fil: Aredes Fernández, Pedro Adrián. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnologico Conicet Noa Sur. Instituto Superior de Investigaciones Biologicas. Grupo de Investigacion y Desarrollo del Noroeste Argentino | Universidad Nacional de Tucuman. Instituto Superior de Investigaciones Biologicas. Grupo de Investigacion y Desarrollo del Noroeste Argentino.; Argentina Fil: Kritsanida, Marina. Universite de Paris, Descartes; Francia Fil: Grougnet, Raphael. Universite de Paris, Descartes; Francia Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina

The antifungal activity of organic extracts from leaves, bark and hearthwood of the timber tree Prosopis nigra was investigated against fungi responsible for soybean late season diseases. Nine extracts were assayed by the disc diffusion test toward Septoria glycines and Cercospora kikuchii. The methanol extract of leaves was the most active with the lowest inhibitory dose of 200 μg disc−1 on both fungi and diameters of growth inhibition zones of 18.5 mm (C. kikuchii) and 20.5 mm (S. glycines). Tryptamine was the main antifungal constituent of the extract. The methanol extract and tryptamine synergized the effect of difenoconazole. Tryptamine also inhibited cercosporin produced by C. kikuchii in 36–96 % when applied at 25−100 μg and 70 % when provided in mixture with the fungicide. Preventive applications of the extract and the mixture on soybean plants strongly reduced disease severity produced by both fungal pathogens. Our results indicate that the leaf methanol extract of P. nigra deserves further research as an additive of azole fungicides applied for the control of C. kikuchii and S. glycines.

Essential oils from aerial parts of

Extracts from aerial parts of medicinal plants from northwest Argentina were screened for antibacterial activity against the phytopathogenic strains namely CECT 124 (Pseudomonas corrugata), CECT 126 (P. syringae pv. tomato), CECT 225 (Erwinia carotovora var. carotovora), CECT 472 (Agrobacterium tumefaciens) and CECT 792 (Xanthomonas campestres pv. vesicatoria). Leaves and stems of Aspidosperma quebracho-blanco, Schinus fasciculatus, S. gracilipes, Amphilophium cynanchoides and Tecoma stans were separately extracted with solvents of increasing polarity to obtain the dichloromethane (fCH2Cl2), ethyl acetate (fEtAc) and methanol extracts (fMeOH), respectively. Among the thirty extracts tested, only fEtAc from leaves and stems of S. fasciculatus reached the IC50 against the five bacterial strains tested (IC50 = 0.9 mg/ml). The fEtAc from the leaves contained kaempferol, quercetin and agathisflavone which had moderate to strong antibacterial activity. This extract and its identified flavonoids showed synergic (CECT 124,126 and 792) or additive effects (CECT 472 and 225) in mixtures with Kocide 3000. Fil: Terán Baptista, Zareath Pamela. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Gómez, Analia de Los Angeles. Universidad Nacional de Tucumán; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Kritsanida, Marina. Universidad Nacional de Tucumán; Argentina Fil: Grougnet, Raphaël. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Mandova, Tsvetelina. Université Paris Descartes; Francia Fil: Aredes Fernández, Pedro Adrián. Universite Paris Descartes; . Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina Fil: Sampietro, Diego Alejandro. Universite Paris Descartes; . Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina

Essential oils from aerial parts of Senecio nutans, Senecio viridis, Tagetes terniflora and Aloysia gratissima were analysed by GC-MS and their antifungal activities were assayed on toxigenic Fusarium and Aspergillus species. Sabinene (27.6 ± 0.1%), α-phellandrene (15.7 ± 0.3%), o-cymene (9.6 ± 0.2%) and β-pinene (6.1 ± 0.2%) in S. nutans, 9,10-dehydrofukinone (92.7 ± 0.2%) in S. viridis, β-thujone (36.1 ± 0.1%), α-thujone (32.2 ± 0.2%), 1,8-cineol (10.7 ± 0.1%) and sabinene (6.2 ± 0.2%) in A. gratissima, and cis-tagetone (33.6 ± 0.2%), cis-β-ocimene (17.1 ± 0.2%), trans-tagetone (17.0 ± 0.1%), cis-ocimenone (8.0 ± 0.2%) and trans-ocimenone (8.2 ± 0.1%) in T. terniflora. The oils showed moderate antifungal activity (1.2 mg/mL > MIC >0.6 mg/mL) on the Fusarium species and a weak effect on Aspergillus species. The antifungal activity was associated on F. verticillioides to the high content of cis-tagetone, trans-tagetone, cis-β-ocimene, cis-ocimenone, trans-ocimenone and on F. graminearum due to the total content of oxygenated sesquiterpenes and 9,10- dehydrofukinone. The oil of S. viridis synergized the effect of fungicides and food preservatives on F. verticillioides.

Aims: The purpose of this research was to identify antifungal compounds from leaves of Schinus and Schinopsis species useful for the control of toxigenic Fusarium species responsible of ear rot diseases. Methods and Results: Leaves of Schinopsis (S. lorentzii and S. haenkeana) and Schinus (S. areira, S. gracilipes and S. fasciculatus) were sequentially extracted with dichloromethane, ethyl acetate and methanol. The antifungal activity of the fraction soluble in methanol of these extracts (fCH2Cl2, fAcEt and fMeOH, respectively) was determined by the broth microdilution method and the discdiffusion method. The minimum inhibitory dose (MID), the diameter of growth inhibition (DGI) and the minimum concentration for 50% inhibition of fungal growth (MIC50) were calculated. The fCH2Cl2 and fAcEt of the Schinopsis species had the lowest MID and MIC50 values and the highest DGI. The antifungal compounds were identified as lupeol and a mix of phenolic lipids. The last one had the highest antifungal activity with MIC50 31– 28 lg g 1 and 165–150 lg g 1 on Fusarium graminearum and Fusarium verticillioides, respectively. The identified metabolites completely inhibited fumonisin and deoxynivalenol production at lower concentrations than ferulic acid, a natural antimycotoxigenic compound. Conclusions: It was proven that lupeol and phenolic lipids were inhibitors of both fungal growth and mycotoxin production of toxigenic Fusarium species. This fact is specially interesting in the control of the toxigenic Fusarium species because several commercial antifungals showed to stimulate mycotoxin biosynthesis at sublethal concentrations. Significance and Impact of the Study: Control of toxigenic Fusarium species requires compounds able to inhibit both fungal growth and mycotoxin production. Our results suggest that the use of lupeol as food preservative and the phenolic lipids as fungal growth inhibitors of F. verticillioides and F. graminearum did not imply an increase in mycotoxin accumulation. Fil: Aristimuño Ficoseco, Maria Eugenia del Milagro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Audenaert, K.. University of Ghent; Bélgica Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina

Fusarium meridionale and F. boothii cause Gibberella Ear Rot (GER) in maize. This study determined the effects of temperature (5–35 °C) and water activity (0.90–0.995 aw) on the growth, and deoxynivalenol (DON) and nivalenol (NIV) production of F. meridionale and F. boothii strains in maize grains. Fusarium graminearum sensu stricto strains from wheat were also tested. The three Fusarium species grew best at 0.995 aw and 25 °C. Growth was absent or marginal at 0.90 aw regardless of temperature. F. meridionale and F. boothii were sensitive to 30 °C and more affected by water stress than F. graminearum sensu stricto. The highest DON levels were at 0.995–0.97 aw and 30 °C and at 0.97 aw and 20 °C for F. graminearum sensu stricto, and at 0.995–0.97 aw and 20 °C for F. boothii. Fusarium meridionale reached maximum NIV accumulation at 0.995 aw and 20 °C. This produced DON at negligible levels compared to the other two Fusarium species. Growth of F. meridionale and F. boothii was well adapted to the usual autumn high humidity and mild temperatures associated with GER in northwest Argentina. Control strategies during grain development should be taken into account to reduce the risk of the presence of DON and NIV in the harvested grains.

A 96-well plate micromethod was developed to measure 5-n-alkylresorcinols (5nARs) in cereal grains and food derived products. The 5nARs reacted in alkaline alcoholic medium with Fast Blue RR ½ZnCl2 salt to yield coloured azo-derivatives. The highest sensitivity for 5nARs was obtained at 490 nm with 0.025% ethanolic Fast Blue RR and 5% K2CO3. This reaction showed good linearity for olivetol (0.05–0.20 μg). Contents of 5nARs determined in cereal grains and derived products by the new Fast Blue RR micromethod were highly correlated (R2 = 0.9944) with those obtained by a Fast Blue B method currently used. A Bland–Altman analysis indicated a small positive bias near to zero (R2 = 0.0401), suggesting that the methods can be interchangeably used. The new reaction is completed in 15 min and the coloured products are read within the 15 min after completion. The micromethod offers a fast analysis of 5nARs in cereal grains and derived products with low consumption of reagents and solvents. Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Jiménez, Cristina Marisol. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Belizán, María Melina Eleonora. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Quimica del Noroeste; Argentina

Twenty six isolates of Fusarium graminearum from grains of maize hybrids harvested in ±west Argentina were grown on autoclaved rice grain to assess their ability to produce type B trichothecenes. Chemical analysis indicated that 38% of isolates were nivalenol (NIV) producers only, 31% were major NIV producers with high DON(deoxynivalenol)/NIV ratios, 8% were major DON producers with minor NIV production, and 23% were DON producers only. Isolates showed a high variability in their toxigenic potential which was not related to fungal biomass. The distribution of the different chemotypes as well as the high and the low trichothecene-producing Fusarium isolates could not be associated to a geographical origin. Our results confirmed for the first time that isolates of Fusarium graminearum from maize of northwest Argentina are able to produce DON and NIV. A substancial contamination with both NIV and DON is likely in maize from northwest Argentina. Their contents should be quantified in regional surveillances for mycotoxin contamination. Fil: Sampietro, Diego Alejandro. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Apud, Gisselle Raquel. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Belizán, María Melina Eleonora. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vattuone, Marta Amelia. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Catalan, Cesar Atilio Nazareno. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Tucumán. Instituto de Quimica del Noroeste; Argentina; Argentina

An antifungal activity-directed fractionation of leaf constituents from Schinopsis balansae on Fusarium graminearum yielded a fraction mainly made of a mixture of four 3-n-heptadec(en)ylcatechols (PALK). The PALK fraction showed on macroconidia germination a MIC100value of 500 μg/mL which was twofold higher than that required for prothioconazole (MIC100 = 250 μg/mL). Sublethal concentrations of PALK modify the morphogenesis in germinating macroconidia, and decreased fungal production of H2O2and deoxynivalenol biosynthesis at early fungal growth. Mixes of PALK and prothioconazole showed a synergic interaction. Our findings suggest that PALK constituents might restrict the adherence of F. graminearum to the surface of its hosts and its virulence on susceptible cereals. They deserve further research as additives of azole fungicides against F. graminearum. Fil: Aristimuño Ficoseco, Maria Eugenia del Milagro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: Sequin, Christian Javier. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ciencias Agropecuarias; Argentina Fil: Aceñolaza, Pablo Gilberto. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Entre Ríos. Facultad de Ciencias Agropecuarias; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: Catalan, Cesar Atilio Nazareno. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Instituto de Química del Noroeste. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Química del Noroeste; Argentina

Propolis is a bee hive product with complex chemical composition that was used as medicine from ancient times. The immunomodulatory activity of partially purified propolis extracts (PPPEs), galangin and pinocembrin was studied through their in vitro effect on neutrophil chemotactic and phagocytic activities. PPPEs (15 samples from each of 10 different beehives) were more effective (around 45% and 50% for a concentration of 40 μg of PPPEs/mL of active substance) as chemotactic agents than galangin and pinocembrin (around 20 and 25%, respectively) and showed higher neutrophil phagocitic activity (270 ± 10% for beehive 10) than galangin and pinocembrin (180 ± 10 and 135 ± 9%, respectively). The highest effect of PPPEs can be attributed to a synergistic effect among components of PPPEs. The results of this study indicated that PPPEs, galangin and pinocembrin stimulate neutrophil chemotactic activity at various concentrations suggesting their possible use for patients suffering neutrophil disfunction. The nitro blue tetrazolium results indicated that the extracts and isolated flavonoids have the capacity to scavenge active radicals suggesting that they would be valuable in the treatment of diseases associated with free radical damage. Consequently, the in vitro results suggest a potential value of these propolis extracts in the medical field. Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina

Essential oils from aerial parts of Acantholippia deserticola, Artemisia proceriformis, Achillea micrantha and Libanotis buchtormensis were analysed by GC–MS. The major compounds identified were β-thujone (66.5 ± 0.2%), and trans-sabinyl acetate (12.1 ± 0.2%) in A. deserticola; α-thujone (66.9 ± 0.4%) in A. proceriformis; 1,8-cineole (26.9 ± 0.5%), and camphor (17.7 ± 0.3%) in A. micrantha and cis-β-ocimene (23.3 ± 0.3%), and trans-β-ocimene (18.4 ± 0.2%) in L. buchtormensis. The oils showed a weak antimicrobial effect (MIC100 > 1.5 mg/ml) on most phytopathogens tested. A moderate antimicrobial activity (MIC100 between 0.5 and 1.5 mg/ml) was displayed by the oils of A. deserticola, A. micrantha and L. buchtormensis on Septoria tritici and by the oil of A. deserticola on Septoria glycine. The antimicrobial activity was associated to the contents of β-thujone, trans-sabinyl acetate and trans-sabinol. Our results indicate that the tested essential oils have little inhibitory potency not suitable for use as plant protection products against the phytopathogens assayed.

The composition of essential oils from leaves of Kazakhstan medicinal plants was analysed by GC-MS. The major compounds identified were 1,8-cineole (34.2%), myrcene (19.1%) and α-pinene (9.4%) in Ajania fruticulosa; 1,8-cineole (21.0%), β-thujone (11.0%), camphor (8.5%), borneol (7.3%) and α-thujone (6.5%), in Achillea nobilis; camphor (47.3%), 1,8-cineole (23.9%), camphene (9.8%) and β-thujone (6.0%) in Artemisia terrae-albae; 1,8-cineole(55.8%) and β-pinene (6.2%) in Hyssopus ambiguus; α-thuyene(46.3%) and δ-cadinene(6.3%) in Juniperus sibirica; sabinene (64%) in Juniperus sabina; and α-pinene (51.5%), β-phellandrene (11.2%) and δ-cadinene (6.3%) in Pinus sibirica. The essential oils did not show antifungal effect (MIC > 1.20 mg/mL) on Aspergillus carbonarius and Aspergillus niger, while the oils from A. nobilis, A. terrae-albae, H. ambiguus and J. sabina exhibited moderate and moderate to weak antimicrobial activities on Fusarium verticillioides (MIC = 0.60 mg/mL) and Fusarium graminearum (MIC = 0.60–1.20 mg/mL), respectively. A principal component analysis associated the antifungal activity (r2 > 0.80, p = 0.05) with the presence of borneol, camphor, camphene, 1,8-cineole,α- and β-thujone, and of the oxygenated monoterpenes.

An α-galactosidase was isolated from a culture filtrate of Lenzites elegans (Spreng.) ex Pat. MB445947 grown on citric pectin as carbon source. It was purified to electrophoretic homogeneity by ammonium sulfate precipitation, gel filtration chromatography and anion-exchange chromatography. The relative molecular mass of the native purified enzyme was 158 kDa determined by gel filtration and it is a homodimer (Mr subunits = 61 kDa). The optimal temperature for enzyme activity was in the range 60-80 °C. This α-galactosidase showed a high thermostability, retaining 94 % of its activity after preincubation at 60 °C for 2 h. The optimal pH for the enzyme was 4.5 and it was stable from pH 3 to 7.5 when the preincubation took place at 60 °C for 2 h. It was active against several α-galactosides such as p-nitrophenyl-α-D-galactopyranoside, α-D-melibiose, raffinose and stachyose. The α-galactosidase is a glycoprotein with 26 % of structural sugars. Galactose was a non-competitive inhibitor with a Ki = 22 mM versus p-nitrophenyl-α-D-galactoside and 12 mM versus α-D-melibiose as substrates. Glucose was a simple competitive inhibitor with a Ki = 10 mM. Cations such as Hg(2+) and p-chloromercuribenzoate were also inhibitors of this activity, suggesting the presence of -SH groups in the active site of the enzyme. On the basis of the sequence of the N-terminus (SPDTIVLDGTNFALN) the studied α-galactosidase would be a member of glycosyl hydrolase family 36 (GH 36). Given the high optimum temperature and heat stability of L. elegans α-galactosidase, this fungus may become a useful source of α-galactosidase production for multiple applications.

The antimicrobial activities and toxicity of infusion, decoction and tincture of Caesalpinia paraguariensis Burk. bark (CPBEs) were investigated to validate its traditional use as drink additive and to identify microbicidal component(s). The minimum inhibitory concentrations (MIC) of CPBEs against aerobic bacteria (Gram-negative and Gram-positive species) were determined using standardised dilution methods. The LC 50 were determined by Brine Shrimp Test. CPBEs showed bacteriostatic and bactericidal activity against tested strains. The highest activity was observed for infusion (MIC:200 μg/mL) against Morganella morganii, Erwinia carotovora, Bacillus spp., Staphylococcus spp. and Enterococcus spp. The bacterial species were susceptible to CPBEs (MIC:200-1993 μg/mL) at lower concentration than sodium benzoate, a known food preservative. Two bioactive components were isolated from liophylised infusion by bio-guided chromatographic procedures; these were identified by spectrometric techniques as ellagic and 3-O-methylellagic acids. This study demonstrated that C. paraguariensis bark infusion it is safe for human consumption and a possible source of food natural preservatives. Fil: Sgariglia, Melina Araceli. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Soberon, Jose Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Quiroga, Emma Nelly. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina

Random S5 inbreds derived from three F2 maize (Zea mays L.) populations (L1934 × LP918, LP915 × LP2541 and L7310 × L7266) were selected for ear rot resistance after inoculation with a low-fumonisin producing isolate belonging to F. proliferatum. The four less susceptible and the four most susceptible inbreds from each population were crossed and F1 seeds were pooled. Resistant and susceptible pools from each population were evaluated for disease severity (percentage of the ear visibly diseased) after inoculation with the isolate used for selection, and high toxigenic isolates belonging to F.verticillioides and F. graminearum. Grain mycotoxin concentration was assessed by ELISA. Differences in disease resistance to each fungus were observed between resistant and susceptible pools in most populations and environments indicating that selection after inoculation with a single species might be effective to develop broad-based resistance to Fusarium. Resistant pools exhibited, after inoculation with F. verticillioides, low grain fumonisin concentrations in most populations and years. Positive genotypic correlations between disease severity and fumonisin concentration (0.89 < rg < 0.98, depending on fungal species and year) indicate that selection for disease severity accounted for most of the variability for field fumonisin accumulation. Selection seemed to be also effective to reduce grain deoxynivalenol and zearalenone concentrations after inoculation with F. graminearum. Ratios between grain deoxynivalenol concentration and disease severity were lower in L7310 × L7266 than those observed in the other populations suggesting that mechanisms affecting mycotoxin accumulation might exist in this population and additional responses should be feasible if including deoxynivalenol concentration as another selection parameter.

Ethnopharmacological relevance: Leaf extracts from Tripodanthus acutifolius (Ruiz and Pavón) Van Tieghem have long been used in argentinean traditional medicine as anti-inflammatory, however, there is no scientific evidence which supports this use in the literature. Aim of the study: The present study was conducted to evaluate the ability of five phenolic compounds purified from infusion prepared from Tripodanthus acutifolius leaves to inhibit key enzymes in inflammatory processes. As anti-inflammatory compounds frequently possess free radical scavenging activities, purified substances were comparatively evaluated to asses their free radical scavenging properties. Genotoxic effects were also evaluated. Materials and Methods: Compounds were evaluated on their ability to inhibit hyaluronidase and cyclooxygenase-2 (COX-2) activities to assess their anti-inflammatory capacities. Free radical scavenging activity was assessed by: 1,1-diphenyl-2-picryl-hydrazyl radical (DPPH), superoxide anion assay and the inhibition on lipid peroxidation. Genotoxicity was evaluated by Bacillus subtilis rec assay. Results: Fractionation of Tripodanthus acutifolius infusion yielded a novel phenylbutanoid derivative (tripodantoside) and four known flavonoid glycosides (rutin, nicotiflorin, hyperoside and isoquercitrin). Flavonoids produced higher inhibition on hyluronidase activity (IC50≈1.7mM) than tripodantoside (IC50=27.90mM). A similar COX-2 inhibition activity was exerted by tripodantoside and monoglycosilated flavonoids (IC50∼50μM). Compounds were strong radical scavengers, with effective concentration 50 (EC50) values for DPPH in the range of 2.7-6.3μg/mL, and for superoxide anion in the range of 3.9-8.7μg/mL. All compounds scavenged peroxyl radicals in the lipid peroxidation assay. The substances showed no genotoxic effects. Conclusions: The anti-inflammatory effects, free radical scavenging activities and lack of genotoxicity of purified compounds may support the folk use of infusion from Tripodanthus acutifolius leaves as anti-inflammatory. Fil: Soberon, Jose Rodolfo. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales; Argentina Fil: Sgariglia, Melina Araceli. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales; Argentina Fil: Sampietro, Diego Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales; Argentina Fil: Quiroga, Emma Nelly. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia. Instituto de Estudios Vegetales; Argentina Fil: Vattuone, Marta Amelia. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán; Argentina. Universidad Nacional de Tucumán. Facultad de Bioquímica, Química y Farmacia; Argentina

Fusarium species are worldwide causal agents of ear rot in cereals. Their toxigenic potential is a health risk for both humans and animals. In Argentina, most identification of these fungi has been based on morphological and cross-fertility criteria which are time consuming and require considerable expertise in Fusarium taxonomy and physiology. DNA based approaches have been reported as rapid, sensitive and specific alternatives to identify the main fumonisin and trichothecene-producing Fusarium species. In this work, we used PCR assays and the partial sequence of TEF1-alpha gene (Translation Elongation Factor-1 alpha) to identify the fumonisin and trichothecene-producing species in Fusarium isolates from diverse regions of Argentina. The relative efficiency and reliability of those methods to improve mycotoxin risk prediction in this country were also assessed. Species-specific PCR assays were targeted toward multicopy IGS (Intergenic Spacer of rDNA units) and on the toxin biosynthetic genes FUM1 (fumonisins) and TRI13 and TRI7 genes (trichothecenes). PCR assays based on FUM1 gene and IGS sequences allowed detection and discrimination of the fumonisin producers Fusarium proliferatum and Fusarium verticillioides. Molecular identification of nonfumonisin producers from Gibberella fujikuroi species complex was possible after determination of TEF1-alplha gene sequences, which indicated the presence of Fusarium subglutinans, Fusarium andiyazi and Fusarium thapsinum. TEF-1 alpha gene sequences also allowed discrimination of the different species of the Fusarium graminearum complex (F. graminearum sensu lato) as F. graminearum sensu stricto, Fusarium meridionale and Fusarium boothii. The last two species belonged to NIV chemotype and were detected for the first time in the subtropical region of Argentina while F. graminearum sensu stricto was DON producer only, which was also confirmed by specific PCR assays based on TRI137/TRI7 genes. Our results indicated that the PCR assays evaluated in this work are reliable diagnostic tools to detect the main toxigenic Fusarium species associated to cereal grains in Argentina. An extensive epidemiological survey based on the approach presented in this work is currently in progress to know the mycotoxigenic hazard of Fusarium species in cereal grains from the subtropical region of Argentina.

The present work describes the purification and characterization of a novel extracellular polygalacturonase, PGase I, produced by Pycnoporus sanguineus when grown on citrus fruit pectin. This substrate gave enhanced enzyme production as compared to sucrose and lactose. PGase I is an exocellular enzyme releasing galacturonic acid as its principal hydrolysis product as determined by TLC and orcinol-sulphuric acid staining. Its capacity to hydrolyze digalacturonate identified PGase I as an exo -polygalacturonase. SDS-PAGE showed that PGase I is an N -glycosidated monomer. The enzyme has a molecular mass of 42 kDa, optimum pH 4.8 and stability between pH 3.8 and 8.0. A temperature optimum was observed at 50–60 °C, with some enzyme activity retained up to 80 °C. Its activation energy was 5.352 cal mol −1 . PGase I showed a higher affinity towards PGA than citric pectin (Km = 0.55 ± 0.02 and 0.72 ± 0.02 mg ml −1 , respectively). Consequently, PGase I is an exo -PGase, EC 3.2.1.82.