The indiscriminate use of herbicides makes weeds resistant, increasing the demand for more efficient herbicides. Novel potential herbicides were prepared from sorbic acid, being 8 dienamides and 6 epoxides. Subsequently, phytotoxic activities of sorbic acid and 14 synthesized compounds were evaluated against sorghum, onion, cucumber, lettuce, and beggartick seeds. Furthermore, the protein target of the most active substance in plants was identified in silico. Among the tested compounds, (2E,4E)-N-(p-fluorophenyl)hexa-2,4-dienamide (2c), (2E,4E)-N -(o-naphthyl) hexa-2,4-dienamide (2d), (E)-N-methylphenyl-3-((2R,3R)-3-methyloxiran-2-yl)acrylamide (3b), (E)-N-bromophenyl-3-((2R,3R)-3-methyloxiran-2-yl)acrylamide (3e), and (E)-N-methoxyphenyl-3-((2R,3R)-3-methyloxiran-2-yl)acrylamide (3f) showed pronounced inhibitory activities, both of shoots and roots, making them candidates for promising herbicides. All compounds altered seed development, and compounds 2c, 2d, 3b, 3e, and 3f presented equal or superior results to the commercial control herbicide S-metolachlor. Docking studies suggest that enzyme mitogen-activated protein kinase can be the target of those compounds. resistant epoxides Subsequently 1 sorghum onion cucumber lettuce seeds Furthermore silico 2E,4ENpfluorophenylhexa2,4dienamide 2E4ENpfluorophenylhexa24dienamide EENpfluorophenylhexadienamide 2E,4E N p fluorophenyl hexa 2,4 dienamide 2E 4E 2 4 E 2c c , (2c) 2E,4EN 2E4EN EEN onaphthyl o naphthyl -(o-naphthyl hexa2,4dienamide hexa24dienamide hexadienamide 2d d (2d) ENmethylphenyl32R,3R3methyloxiran2ylacrylamide ENmethylphenyl32R3R3methyloxiran2ylacrylamide ENmethylphenylRRmethyloxiranylacrylamide methylphenyl 3 2R,3R methyloxiran yl acrylamide 2R 3R R 3b b (3b) ENbromophenyl32R,3R3methyloxiran2ylacrylamide ENbromophenyl32R3R3methyloxiran2ylacrylamide ENbromophenylRRmethyloxiranylacrylamide bromophenyl 3e e (3e) ENmethoxyphenyl32R,3R3methyloxiran2ylacrylamide ENmethoxyphenyl32R3R3methyloxiran2ylacrylamide ENmethoxyphenylRRmethyloxiranylacrylamide methoxyphenyl f (3f roots development Smetolachlor. Smetolachlor S metolachlor. metolachlor S-metolachlor mitogenactivated mitogen activated 4ENpfluorophenylhexa2 4dienamide ENpfluorophenylhexa 2E4E EE 24 2, (2c 4EN EN hexa2 (2d ENmethylphenyl32R 3R3methyloxiran2ylacrylamide ENmethylphenyl ylacrylamide 2R3R RR (3b ENbromophenyl32R ENbromophenyl (3e ENmethoxyphenyl32R ENmethoxyphenyl 4ENpfluorophenylhexa ENmethylphenylR Rmethyloxiranylacrylamide ENbromophenylR ENmethoxyphenylR

Termites are insects with a complex social organization on castes among which soldiers are the only responsible for the colony defense. The soldiers of the Nasutitermes genus use chemical defense, which comprises a mixture of mono, sesqui and mainly diterpenes. The new diterpene 2b,3a-dihydroxy-trinervita-1(15),11-diene, found in Nasutitermes macrocephalus, was isolated using preparative high performance liquid chromatography (HPLC) and identified by gas chromatography-mass spectrometry (GC-MS) and nuclear magnetic resonance (NMR) techniques. The absolute stereochemistry determined was supported by ab initio calculations and by comparison of the experimental circular dichroism data with the theoretical results. The isolated diterpene was evaluated against the bacteria Staphylococcus aureus (Gram-positive) and Escherichia coli (Gram-negative), and the fungi Candida albicans and Aspergillus brasiliensis. The isolated diterpene presented minimal inhibitory concentrations of 62.5 and 62.5 µg mL-1 against S. aureus and C. albicans, respectively.

To contribute to the development of new products to control the coffee leaf miner (Leucoptera coffeella), which is a major pest for coffee plants, this work aimed to select plant species able to produce substances active against this insect and to identify the active substances in the selected plant and their protein targets in the insect. Among the extracts of nineteen plant species, only that from leaves of Merremia tomentosa (Choisy) Hall. f. (Convolvulaceae) reduced the oviposition of L. coffeella on leaves of coffee plants. This extract was submitted to successive fractionation steps to achieve the isolation and identification of two active substances, ursolic acid (UA) and cis-tiliroside (CT). An in silico study showed that UA inhibits glycogen phosphorylases (GP) by binding to their allosteric site, while CT probably inhibits xanthine dehydrogenases (XT). As both GP and XT appear to be essential enzymes for insects, these results suggest that, during evolution, L. coffeella learned to identify plants producing inhibitors of these enzymes to avoid oviposition on such plants. Thus, both UA and CT have potential to be used as lead compounds in the development of new products for the control of the coffee leaf miner.

In a previous study, substances with nematicidal properties were detected in the bark of Cryptocarya aschersoniana. Continuing such study, the methanol extract from this plant underwent fractionation guided by in vitro assays with the plant-parasitic nematode Meloidogyne exigua. Two active compounds were isolated and identified by spectroscopic methods as (E)-6-styrylpyran-2-one and (R)-goniothalamin. The latter compound was also active againstMeloidogyne incognita. In silico studies carried out with (R)-goniothalamin and the enzyme fumarate hydratase, which was extracted from the genome of Meloidogyne hapla and modeled using computational methods, suggested that this substance acts against nematodes by binding to a cavity close to the active site of the enzyme.

Com o objetivo de contribuir para o desenvolvimento de produtos para o controle de Meloidogyne exigua, as bactérias Bacillus cereus e B. subtilis foram cultivadas em meio líquido de cultura para produzirem metabólitos ativos contra este nematoide parasita de plantas. Os fracionamentos dos extratos em diclorometano dos meios de cultura produziram uracila, 9H-purina e di-idrouracila. Todos os compostos foram ativos contra M. exigua, sendo o último o mais eficiente. Ele apresentou CL50 de 204 µg/mL contra o nematoide, enquanto uma CL50 de 260 µg/mL foi observada para o nematicida comercial carbofuran. Uma busca por complexos proteína-ligante nos quais o ligante fosse estruturalmente similar à di-idrouracila resultou na seleção de fosforibosiltransferases, cujas sequências foram utilizadas em uma busca in silico no genoma de M. incognita por sequência de aminoácidos semelhante. A sequência resultante foi modelada e di-idrouracila e 9H-purina foram inseridos nos sítios ativos desta provável fosforibosiltransferase, resultando em complexos proteína-ligante que foram submetidos a simulações por dinâmica molecular. Cálculos das energias livres de ligação destes complexos revelaram que a constante de dissociação de di-idrouracila e 9H-purina da enzima é da ordem de 8,3 x 10-7 e 1,6 x 10-6 M, respectivamente. Consequentemente, estas substâncias e a provável fosforibosiltransferase podem ser de grande utilidade para o desenvolvimento de novos produtos para o controle de M. exigua.

To contribute to the development of products to controlMeloidogyne exigua, the bacteria Bacillus cereus and B. subtilis were cultivated in liquid medium to produce metabolites active against this plant-parasitic nematode. Fractionation of the crude dichloromethane extracts obtained from the cultures afforded uracil, 9H-purine and dihydrouracil. All compounds were active against M. exigua, the latter being the most efficient. This substance presented a LC50 of 204 µg/mL against the nematode, while a LC50 of 260 µg/mL was observed for the commercial nematicide carbofuran. A search for protein-ligand complexes in which the ligands were structurally similar to dihydrouracil resulted in the selection of phosphoribosyltransferases, the sequences of which were used in an in silico search in the genome of M. incognita for a similar sequence of amino acids. The resulting sequence was modelled and dihydrouracil and 9H-purine were inserted in the active site of this putative phosphoribosyltransferase resulting in protein-ligand complexes that underwent molecular dynamics simulations. Calculation of the binding free-energies of these complexes revealed that the dissociation constant of dihydrouracil and 9H-purine to this protein is around 8.3 x 10-7 and 1.6 x 10-6 M, respectively. Consequently, these substances and the putative phosphoribosyltransferase are promising for the development of new products to control M. exigua.

Meloidogyne exigua is a plant-parasitic nematode that causes great losses to coffee farmers. Thus, to contribute to the development of new products to control this parasite, the present work studied the effect of volatile organic compounds (VOC) on the nematode and coffee plants (Coffea arabica), since these compounds are known to be used in plant defense against other agronomical pests. The number of galls of M. exigua was reduced when the aerial part of coffee plants was sprayed with combinations of methyl jasmonate + jasmone, (Z)-hex-3-en-1-ol + (E)-hex-3-en-1-ol + (Z)-hex-2-en-1-ol + (E)-hex-2-enal, (E)-hex-2-enyl acetate + hexyl acetate, meso-butane-2,3-diol + butane-1,2-diol + butane-1,3-diol + butane-1,4-diol, 3-hydroxybutan-2-one + 4-hydroxybutan-2-one or linalyl acetate + nerolidol. These VOC also caused alterations in the concentrations of substances such as alkaloids, phenols, amino acids and carbohydrates, in the roots of coffee plants. The findings suggest that these VOC may be explored as potential products for the control of M. exigua in coffee plants.

Para contribuir para o desenvolvimento de produtos antibacterianos obtidos de própolis produzidos por abelhas nativas do Brasil, 29 tipos de própolis coletados de diferentes colméias no Estado de Minas Gerais, Brasil, foram avaliados quanto à atividade in vitro contra Aeromonas hydrophila, Bacillus subtilis, Pseudomonas aeruginosa, e Staphylococcus aureus. Dentre as amostras de abelhas nativas, somente a de Frieseomelitta varia (Lepeletier, 1836) inibiu, em teste in vitro, o crescimento bacteriano. Consequentemente, essa própolis foi submetida a métodos cromatográficos para o fracionamento biomonitorado por teste de difusão em Agar com as referidas bactérias, o que resultou no isolamento e identificação do ácido 3,5-diprenil-4-hidroxicinâmico (artepelin C), que apresentou CIM de 62,5 e 250 µg/mL frente a B. subtilis e S. aureus, respectivamente. Esses resultados indicam o potencial de F. varia para a produção de própolis terapêutica.

To contribute to the development of antibacterial products from propolis produced by native Brazilian bees, twenty-nine samples of propolis collected from hives in the state of Minas Gerais, Brazil, were screened for in vitro activity against Aeromonas hydrophila, Bacillus subtilis, Pseudomonas aeruginosa, and Staphylococcus aureus. Among the samples from native Brazilian bees, only that from Frieseomelitta varia (Lepeletier, 1836) inhibited in vitro bacterial growth. Consequently, this propolis underwent fractionation by chromatographic methods monitored through Agar-diffusion assays with these bacteria, which resulted in the isolation and identification of 3,5-diprenyl-4-hydroxycinnamic acid (artepillin C), which showed MIC of 62.5 and 250 µg/mL against B. subtilis and S. aureus, respectively. This result indicates the potential of F. varia to produce therapeutic propolis.

Com o objetivo de selecionar rizobactérias produtoras de fitotoxinas para uso no controle de plantas daninhas, vinte e cinco isolados bacterianos previamente obtidos das rizosferas de diferentes plantas foram cultivados em meio líquido e, após remoção das células por centrifugação, as fases líquidas foram liofilizadas e os resíduos obtidos foram submetidos à extração com acetato de etila/metanol. Os extratos foram concentrados sob vácuo até secura e dissolvidos em água e solução de sacarose para serem submetidos a testes in vitro de germinação de sementes de alface (Lactuca sativa L.) e de crescimento de coleóptilos de trigo (Triticum aestivum L.). Embora a maior parte das amostras tenha desfavorecido o crescimento dos coleóptilos de trigo, somente as provenientes de quatro isolados reduziram a germinação das sementes de alface. Dois isolados de Bacillus cereus, um isolado de B. pumilus e um de Stenotrophomonas maltophilia foram os microrganismos mais promissores para a produção de fitotoxinas, com possibilidade de uso no desenvolvimento de novos produtos para o controle de plantas daninhas.

In order to select phytotoxin producing rhizobacteria to control weed plants, twenty five bacterial strains previously isolated from the rhizospheres of various plants were grown in a liquid medium and, after cell removal by centrifugation, the liquid phases were freeze-dried and the products were extracted with ethyl acetate/methanol. The extracts were concentrated to dryness under vacuum and dissolved in water and sucrose solution to be submitted to in vitro assays of lettuce (Lactuca sativa L.) seed germination and wheat (Triticum aestivum L.) coleoptile growth. Although most samples affected coleoptile growth, only those from four strains reduced lettuce seed germination. Two strains of Bacillus cereus, one strain of B. pumilus and one of Stenotrophoonas altophilia were the most promising microorganisms for producing phytotoxin and, consequently, for the development of new weed control products.

Com vistas a descobrir antimicrobianos de origem natural para uso terapêutico ou para a preservação de alimentos, o extrato metanólico das partes aéreas de Solanum palinacanthum foi submetido a fracionamentos direcionados por testes para avaliar a atividade antibacteriana e antifúngica. Em decorrência, o flavonóide rutina e o ácido 3,5-dicafeoilquínico foram isolados por cromatografia em coluna e por cromatografia líquida de alta eficiência, para serem identificados por espectrometria de massas e de ressonância magnética nuclear. As concentrações inibitórias mínimas (CIM) do derivado do ácido cafeico contra Aeromonas hydrophila, Bacillus subtilis, Staphylococcus aureus e o fungo Aspergillus ochraceus foram 250, 1000, 1000 e > 568µg/mL, respectivamente. Contra os mesmos organismos, os valores de CIM para a rutina foram 1000, > 1000, > 1000 e 35µg/mL, respectivamente. A rutina mostrou-se muito promissora para o controle de A. ochraceus, pois seu valor de CIM contra tal fungo foi bem próximo ao observado para o cloreto de benzalcônio, que é empregado como fungicida no Brasil.

To find out natural antimicrobial agents as alternative in therapeutics and to preserve food, the methanol extract of Solanum palinacanthum aerial parts was submitted to purification steps guided by antibacterial and antifungal assays. As a consequence, the flavonoid rutin and 3,5-dicaffeoylquinic acid were isolated by column chromatographyand high performance liquid chromatography, and identified by mass and nuclear magnetic resonance spectrometry. Minimal inhibitory concentrations (MIC) of the quinic acid derivative against Aeromonas hydrophila, Bacillus subtilis, Staphylococcus aureus and the fungus Aspergillus ochraceus were 250, 1000, 1000 and > 568µg/mL, respectively. Against the same microorganisms, MIC for rutin were 1000, > 1000, > 1000 and 35µg/mL, respectively. Rutinwas very promising for A. ochraceus control, since its MIC against such fungus was close to the one observed for benzalkonium chloride, which is used as a fungicide in Brazil.

Observou-se, em estudo preliminar, que o extrato metanólico das folhas de L. lilacina, coletadas no município de Lavras (MG, Brasil), apresentava atividade antibacteriana. Em decorrência, buscou-se purificar e identificar a substância responsável por tal efeito, através de fracionamento do referido extrato direcionado por testes de difusão em agar com Aeromonas hydrophila, Bacillus subtilis, Pseudomonas aeruginosa e Staphylococcus aureus. Após partições com solventes e vários processos cromatográficos, isolou-se o [β-3,4-diidroxifenil)etil]-(3'-O-α-L-ramnopiranosil)-(4'- O-cafeoil)-β-D-glicopiranosídeo, que é conhecido como acteosídeo. A concentração inibitória mínima e a concentração bactericida mínima desta substância para A. hydrophila, B. subtilis, P. aeruginosa e S. aureus foram de 0,12, 1,00, 1,00 e 0,25 mg/mL, respectivamente.

Since the methanol extract of Lantana lilacina leaves collected in the city of Lavras (MG, Brazil) showed antibacterial properties in a preliminary study, a fractionation process guided by agar diffusion assays with Aeromonas hydrophila, Bacillus subtilis, Pseudomonas aeruginosa and Staphylococcus aureus was carried out to purify and identify the active compounds. After solvent partition and several chromatographic steps, [ß-3,4-dihydroxyphenyl)-ethyl]-(3'-O-α-L-rhamnopyranosyl)-(4'- O-cafeoyl)-β-D-glycopyranoside, known as acteoside, was isolated. The minimal inhibition concentration and the minimal bactericidal concentration of such substance against A. hydrophila, B. subtilis, P. aeruginosa and S. aureus were 0.12, 1.00, 1.00 and 0.25 mg/mL, respectively.

Com vistas a contribuir para o desenvolvimento de métodos não agressivos ao meio ambiente, para o controle de plantas invasoras, buscou-se selecionar rizobactérias produtoras de substâncias fitotóxicas. Inicialmente, 35 culturas previamente isoladas de plantas da região sul do Estado de Minas Gerais (Brasil) foram cultivadas em caldo soja tripticaseína. Após remoção das células bacterianas, os líquidos resultantes foram liofilizados e extraídos com metanol/acetato de etila (1:1). Os extratos foram concentrados sob vácuo e submetidos a testes com sementes de alface (Lactuca sativa L.). Os metabólitos produzidos por cinco isolados bacterianos reduziram o número de plântulas normais para valores estatisticamente inferiores aos observados para o controle negativo, sendo os mais expressivos resultados obtidos com Bacillus cereus Frankland and Frankland, isolado de Ricinus communis L., que causou necroses nas radículas de 82,4% das plântulas. Os metabólitos dessa bactéria também impediram a germinação de 52% das sementes de Brachiaria decumbens Stapf e fizeram com que as 48% restantes dessem origem a plântulas anormais. Os metabólitos de B. cereus foram submetidos a processos de purificação direcionados por testes com sementes de alface. Em decorrência, isolou-se uma substância, que será identificada em estudos futuros. Na concentração de 0,057 g/L, causou necrose nas radículas de todas as plântulas de alface provenientes do teste com sementes.

To contribute for the development of environmental friendly methods for weed control, a selection of rhizobacteria able to produce phytotoxic substances was carried out. Initially, 35 strains previously isolated from plants in the south of Minas Gerais State (Brazil) were grown in tryptic soy broth. After removal of bacterial cells, the resulting liquids were freeze-dried and extracted with methanol/ethyl acetate (1:1). The extracts were concentrated under vacuum and dissolved in water to be submitted to a lettuce (Lactuca sativa L.) seed assay. Metabolites produced by five strains reduced the number of normal seedlings to values statistically bellow the one observed for the negative control, being the most expressive results obtained with Bacillus cereus Frankland and Frankland, isolated from Ricinus communis L., which was able to cause rotted rootlets to 82.4% of seedlings. The bacterium metabolites also avoided germination of 52% Brachiaria decumbens Stapf seeds and the remaining 48% resulted in abnormal seedlings. Metabolites from B. cereus were submitted to a purification process guided by the lettuce seed assay. As a consequence, one substance causing rotted rootlets to all lettuce seedlings during the seed assay at 0.057 g/L was isolated and will be identified in future studies.

Inicialmente buscou-se identificar plantas com extratos metanólicos ativos in vitro contra o nematóide Panagrellus redivivus. Das 24 espécies vegetais estudadas, constatou-se que os melhores resultados foram obtidos com Leucaena leucocephala, que causou mortalidade de 93% dos indivíduos expostos ao seu extrato por 24 h. Em seguida, submeteu-se o referido extrato a fracionamento direcionado pelos testes in vitro com P. redivivus, o que possibilitou isolar uma substância nematicida que, segundo análises preliminares, correspondia a um alcalóide.

Initially this work was aimed to identify plant methanolic extracts toxic in vitro to the nematode Panagrellus redivivus. Among 24 plant species studied, Leucaena leucocephala afforded the best results, as 93% of the nematodes were dead after exposure to its extract during 24 h. Thus, such extract was submitted to a purification process guided by in vitro assays with P. redivivus. As a result, one substance active against such nematode was isolated. According to preliminary analysis, that seems to be an alkaloid.

Objetivando o desenvolvimento de novas metodologias de controle de fitonematóides, este trabalho buscou purificar as substâncias nematicidas produzidas por Cunninghamella elegans, Fusarium sp., Paecilomyces lilacinus eP. variotii. Esses fungos foram cultivados em meio líquido Czapek-Dox durante 15 dias, a 25 ºC, em agitador orbital. Em seguida, filtraram-se as misturas, o que permitiu a obtenção de soluções que foram concentradas sob vácuo e submetidas à purificação direcionada por testes in vitro com Meloidogyne incognita. Observou-se que os filtrados de P. lilacinus e P. variotii perdiam suas atividades nematicidas após a concentração sob vácuo, sugerindo que as substâncias ativas produzidas por esses fungos são consideravelmente voláteis. Para o filtrado de Fusarium sp., observou-se perda total da atividade contra M. incognita após fracionamento em coluna de sílica gel, indicando instabilidade da substância nematicida frente às condições empregadas.Do filtrado de C. elegans isolou-se uma substância que, em solução aquosa na concentração de 250 ppm, imobilizou 94% dos juvenis do segundo estádio de M. incognita expostos a tal solução durante 48 h.

In a search for new molecules toxic to plant parasitic nematodes, procedures were set up to purify nematode toxic molecules produced by Cunninghamella elegans, Fusarium sp., Paecilomyces lilacinus and P. variotii. Those fungi were grown in Czapek-Dox liquid medium during 15 days, at 25 ºC, in an orbital shaker. After fungus mycelium removal by filtration, the solutions were concentrated under vacuum and submitted to purification, guided by in vitro assays with Meloidogyne incognita juveniles. As a result, it was observed that P. lilacinus and P.variotii filtrates lost their activity after vacuum concentration, which suggests that the nematocides produced by those fungi are considerably volatile. In the case of Fusarium sp. filtrate, the active substance seems to be unstable, as the activity against M. incognita juveniles completely disappeared after chromatography in a silica gel column. The C. elegans filtrate rendered one isolated substance that immobilized 94% of the M. incognita juveniles exposed during 48 h to an aqueous solution of the substance at 250 ppm.

Visando à obtenção de novas moléculas nematicidas, 18 filtrados de espécies fúngicas e extratos de sete micélios fúngicos foram estudados in vitro quanto aos efeitos sobre a eclosão, motilidade e mortalidade de juvenis do segundo estádio (J2) de Meloidogyne incognita. Filtrados fúngicos foram obtidos de culturas em meio Czapek. Metanol foi usado como solvente para obter extratos dos micélios fúngicos. Filtrados de Paecilomyces lilacinus, Fusarium moniliforme e Fusarium oxysporum reduziram a motilidade e eclosão, e aumentaram a mortalidade (P < 0,05) de J2 de M. incognita de forma semelhante à observada com Aldicarbe. Alguns filtrados fúngicos reduziram a motilidade, mas não causaram a morte de J2. Já os filtrados de Aspergillus flavus, Cylindrocarpon magnusianum, Fusarium solani e Mortierella sp. reduziram apenas a eclosão de J2. Não houve correlação entre a produção de metabólitos tóxicos e a quantidade de micélio fúngico produzido. No caso específico de P. lilacinus, observou-se que após o primeiro dia de cultivo o filtrado fúngico correspondente já apresentava toxidez contra M. incognita. Com o aumento do número de dias de cultivo foi elevado o efeito do filtrado sobre o nematóide, sendo que apenas após 13 dias a mortalidade de J2 alcançou o índice de 100%.

In the search for new nematicidal molecules, 18 fungal species filtrates and seven fungus mycelium extracts were studied in vitro to see their influence on hatching, mobility and mortality of second stage juveniles (J2) of Meloidogyne incognita. Fungal filtrates were obtained from fifteen day Czapek cultures. Methanol was used as solvent to obtain fungus mycelium extracts. Paecilomyces lilacinus, Fusarium moniliforme and Fusarium oxysporum filtrates reduced J2 motility and hatching, and increased J2 mortality (P < 0,05) similarly to Aldicarb. Some fungus filtrates reduced J2 mobility without leading to death. Aspergillus flavus, Cylindrocarpon magnusianum, Fusarium solani and Mortierella sp. filtrates reduced (P < 0,05) only J2 hatching. Production of toxic fungus metabolites was not dependent on the amount of mycelium produced. P. lilacinus produced toxic filtrates after one day culturing. Since then, toxicity increased steadily, reaching 100% J2 mortality on the thirteenth day.