ABSTRACT Essential oils used as bioherbicides are a promising alternative in agriculture. This study aimed to assess the phytotoxic potential of Piper marginatum essential oil through an in vitro study with detached and post-emergence leaves of three commercial species (lettuce, pepper, and tomato) and a weed (Spermacoce verticillata). The experiments were conducted in a randomized design with six treatments that consisted of four concentrations of essential oil (0.001, 0.01, 0.1, and 1% [v/v]) and two control treatments (distilled water, and 1% polysorbate 80 in distilled water). The P. marginatum essential oil had a phytotoxic effect on the leaves of the experimental species. The highest essential oil concentration resulted in greater toxicity effects on tomato, lettuce, and pepper leaves. The 0.1% essential oil concentration completely destroyed the S. verticillata leaves. Seven days after application, the P. marginatum essential oil did not show adverse effects on plant growth in the post-emergence phase. It is recommended to investigate its possible phytotoxic activity in other phases of plant development to understand its mode of action and potential as an alternative to developing bioproducts for weed control. agriculture postemergence post emergence lettuce (lettuce tomato Spermacoce verticillata. . verticillata) 0.001, 0001 0 001 (0.001 01 0.01 1 0.1 v/v vv v [v/v] water 8 water. water) P S application phase 0.001 000 00 (0.00 0.0 0. [v/v 0.00 (0.0 (0. (0 (

RESUMO Óleos essenciais usados como bioherbicidas são uma alternativa promissora na agricultura. O objetivo deste trabalho foi avaliar o potencial fitotóxico do óleo essencial de Piper marginatum, através de estudo in vitro com folhas destacadas e em pós-emergência de três espécies comerciais (alface, pimentão e tomate) e uma planta daninha (Spermacoce verticillata). Os experimentos foram conduzidos em delineamento casualizado com seis tratamentos que consistiram em quatro concentrações de óleo essencial (0,001; 0,01; 0,1 e 1% [v/v]), e dois tratamentos controle (água destilada, e 1% de polissorbato 80 em água destilada). O óleo essencial de P. marginatum apresentou efeito fitotóxico sobre as folhas das espécies experimentais. A maior concentração de óleo essencial resultou em maiores efeitos de toxicidade nas folhas de tomate, alface e pimentão. A concentração de 0,1% de óleo essencial foi suficiente para destruir completamente as folhas de S. verticillata. Sete dias após a aplicação, o óleo essencial de P. marginatum não evidenciou efeitos adversos no crescimento das plantas na fase pós-emergencial. Recomenda-se investigar sua possível atividade fitotóxica em outras fases do desenvolvimento vegetal, para entender o seu modo de ação e potencial como alternativa no desenvolvimento de bioprodutos para controle de plantas daninhas. agricultura pósemergência pós emergência alface, (alface tomate Spermacoce verticillata . verticillata) 0,001 0001 0 001 (0,001 0,01 01 1 0, v/v, vv v/v , v [v/v]) destilada 8 destilada. destilada) P experimentais S aplicação pósemergencial. pósemergencial emergencial. emergencial pós-emergencial Recomendase Recomenda se vegetal daninhas 0,00 000 00 (0,00 0,0 [v/v] (0,0 [v/v (0, (0 (

Resumo A leishmaniose cutânea (LC) é considerada um problema de saúde pública. Os tratamentos atuais apresentam desvantagens por serem invasivos e possuem efeitos colaterais graves, levando a pesquisa de novas alternativas farmacológicas mais eficazes. As plantas são fontes promissoras de bioativos, e por meio de reações químicas pode-se obter novos análogos. O presente estudo objetivou avaliar os efeitos antileishmania do análogo éter n-butil do dilapiol (EBD) extraído das folhas de Piper aduncum. Foi avaliado o potencial citotóxico do EBD nas concentrações de 15,62 a 500 µM em macrófagos peritoneais por 48 h, e em macrófagos RAW 264.7 por 72 h utilizando o método dose-resposta. A atividade antileishmania em promastigotas de L. amazonensis usou-se concentrações de 0,2 a 4,5 µM durante 24, 48 e 72 h e a quantificação da taxa de infecção celular usou-se uma concentração de 4,5 µM do EBD contra as formas amastigotas interiorizado em macrófagos por 24 h e 48 h. O óxido nítrico foi quantificado a partir de macrófagos tratados previamente com o EBD durante 24 h e 48 h. A dosagem das espécies reativas de oxigênio utilizou-se a concentração de 4,5 µM do EBD incubado junto ao acetato de diclorofluoresceína por 1, 3, 6, e 24 h. Para a modelagem molecular do EBD, usou-se a proteína de origem de Leishmania, disponível no banco de dados “Protein Data Bank”. A molécula do estudo não foi tóxica para as células, apresentou uma CC50 de 413 µM em macrófagos peritoneais e de 373,5 µM em RAW 264.7. O análogo inibiu as formas promastigotas de L. amazonensis com uma CI50 de 1,6 µM por 72 horas. O EBD apresentou uma taxa de infecção de 17% e 12%, dilapiol de 24% e 14% e Pentacarinat® de 10% e 9% durante 48 h. O EBD demonstrou uma ligação de energia de -7,8 pelo sítio de ação da enzima U53. Conclui-se que o análogo mostrou atividade antileishmania promissora para futuros testes in vivo. LC (LC pública graves eficazes bioativos podese pode se análogos nbutil n butil (EBD aduncum 1562 15 62 15,6 50 4 2647 264 7 264. doseresposta. doseresposta dose resposta. resposta dose-resposta L usouse usou 02 0 2 0, 45 5 4, utilizouse utilizou 1 3 6 Leishmania Protein Bank. Bank . Bank” células CC CC5 41 3735 373 373, CI CI5 16 horas 17 12 12% 14 Pentacarinat 10 9 7,8 78 8 -7, U53 U Concluise Conclui vivo 156 15, 26 37 7, -7 U5 -

Abstract Cutaneous leishmaniasis (CL) is considered a public health problem. Current treatments have disadvantages because they are invasive and have serious side effects, and thus there is a need for research into new, more effective pharmacological alternatives. Plants are promising sources of bioactive substances, and new analogues can be obtained through chemical reactions. The present study aimed to evaluate the antileishmanial effects of the analog dillapiole n-butyl ether (DBE) extracted from Piper aduncum leaves. The cytotoxic potential of DBE was evaluated at concentrations of 15.62 to 500 µM in peritoneal macrophages for 48 h, and in RAW 264.7 macrophages for 72 h using a dose-response method. The antileishmanial activity in L. amazonensis promastigotes used concentrations of 0.2 to 4.5 µM for 24, 48 and 72 h and the quantification of the cellular infection rate used a concentration of 4.5 µM of DBE against the amastigote forms internalized in macrophages for 24 h and 48 h. Nitric oxide was quantified from macrophages previously treated with DBE for 24 h and 48 h. The dosage of reactive oxygen species used a concentration of 4.5 µM of DBE incubated together with dichlorofluorescein acetate for 1, 3, 6, and 24 h. For the molecular modeling of DBE, the Leishmania protein, available in the “Protein Data Bank” database, was used. The studied molecule was not toxic to cells and presented a CC50 of 413 µM in peritoneal macrophages and 373.5 µM in RAW 264.7. The analogue inhibited promastigote forms of L. amazonensis with an IC50 of 1.6 µM for 72 h. DBE presented an infection rate of 17% and 12%, dillapiole of 24% and 14% and Pentacarinat® of 10% and 9% over 48 h. DBE demonstrated a binding energy of -7.8 for the U53 enzyme. It is concluded that the analogue showed promising antileishmanial activity for future in vivo tests. CL (CL problem alternatives substances reactions nbutyl n butyl (DBE leaves 1562 15 62 15.6 50 4 2647 264 7 264. doseresponse dose response method L 02 0 2 0. 45 5 4. 1 3 6 protein Protein Bank database CC CC5 41 3735 373 373. IC IC5 16 1. 17 12 12% 14 Pentacarinat 10 9 7.8 78 8 -7. U U5 enzyme tests 156 15. 26 37 7. -7 -

Abstract In the community of Santiago Ecatlán, Jonotla, Puebla, located in a cultural region inhabited by Totonacos, the residents take advantage of various plant species with different uses. Among the edible species, some of the Begonia genus (Begoniaceae) stand out, valued mainly for their cultural importance. The petioles of these begonias are consumed as "quelites" and are appreciated both for their flavor and for the economic income they generate through their sale. Factors such as cultural erosion, climate change, the exchange of species between communities from different states, in addition to advances in taxonomic studies, can modify the existing information from previous works about plants. Therefore, the objective of this work was to update the information on edible begonias, their propagation and form of consumption in the community of Ecatlán, through an ethnobotanical study. The information was collected through participant observation, open and semi-structured interviews, and the collection of botanical samples. The following species were identified: Begonia thiemei and B. manicata as the most used, B. heracleifolia with two variants, a probable hybrid between B. heracleifolia and B. manicata, which had possibly been reported by other authors as B. barkeri or B. aff. barkeri and another identified only to the genus, whose morphological characteristics suggest that it could be a new species for science. Regarding the species mentioned by other authors as edible in the area, there was agreement on the use of Begonia heracleifolia, B. manicata and the probable hybrid mentioned above. However, no evidence was found of the presence and/or use of B. fusca, B. incarnata and B. glabra, reported in previous works. Finally, it was recorded that B. nelumbonifolia is not appreciated as food by the local inhabitants. The identified species were found in family gardens, coffee plantations, "ranches" and along roads. Based on the information collected, it was found that the propagation of begonias is carried out in two ways: 1) through the rhizomes and 2) in the case of B. thiemei, by cutting the mature leaf, approximately 2 -3 cm below the leaf blade, and its subsequent planting in soil. These species are available throughout the year, and the preparation of the traditional dish includes various ingredients, including species from the Apiaceae and Piperaceae families.

Resumen En la comunidad de Santiago Ecatlán, Jonotla, Puebla, ubicada en una región cultural habitada por totonacos, los pobladores aprovechan diversas especies vegetales con distintos usos. Entre las especies comestibles, destacan algunas del género Begonia (Begoniaceae), valoradas principalmente por su importancia cultural. Los peciolos de estas begonias se consumen como "quelites" y son apreciados tanto por su sabor como por los ingresos económicos que generan a través de su venta. Factores como la erosión cultural, el cambio climático, el intercambio de especies entre comunidades de diferentes estados, además de los avances en los estudios taxonómicos, pueden modificar la información existente de trabajos anteriores acerca de las plantas. Por lo anterior, el objetivo de este trabajo fue actualizar la información sobre las begonias comestibles, su propagación y forma de consumo en la comunidad de Ecatlán, a través de un estudio etnobotánico. La información se recopiló mediante la observación participante, entrevistas abiertas y semiestructuradas y la recolección de muestras botánicas. Se identificaron las siguientes especies: Begonia thiemei y B. manicata como las más utilizadas, B. heracleifolia con dos variantes, un probable híbrido entre B. heracleifolia y B. manicata, que posiblemente había sido reportado por otros autores como B. barkeri o B. aff. barkeri y otra identificada solamente a género, cuyas características morfológicas hacen suponer que podría ser una especie nueva para la ciencia. En cuanto a las especies mencionadas por otros autores como comestibles en la zona, se coincidió en el uso de Begonia heracleifolia, B. manicata y el probable híbrido mencionado anteriormente. Sin embargo, no se encontraron indicios de la presencia y/o uso de B. fusca, B. incarnata y B. glabra, reportadas en trabajos anteriores. Por último, se registró que B. nelumbonifolia no es apreciada como alimento por los habitantes de la localidad. Las especies identificadas se encontraron en huertos familiares, cafetales, "ranchos" y a lo largo de los caminos. Con base en la información recopilada se encontró que la propagación de las begonias se lleva a cabo de dos formas: 1) a través de los rizomas y 2) en el caso de B. thiemei, mediante el corte de la hoja madura, aproximadamente 2-3 cm por debajo de la lámina foliar, y su posterior plantación en tierra. Estas especies están disponibles durante todo el año, y la preparación del platillo tradicional incluye diversos ingredientes, entre ellos, especies de las familias Apiaceae y Piperaceae.

Abstract This study investigated for the first time the effectiveness of therapeutic baths with essential oil (EO) of Piper hispidum against monogeneans Anacanthorus spathulatus, Notozothecium janauachensis, Mymarothecium boegeri and Linguadactyloides brinkmanni from the gills of Colossoma macropomum, as well as the hematological and histological effects on this fish. In therapeutic baths, 100 mg/L of P. hispidum essential oil and two control groups (water from the culture tank and water from the culture tank with 70% alcohol) were exposed for 1 h/day, with intervals of 48 hours for 3 days, and three replicates each were used. Therapeutic baths with 100 mg/L of P. hispidum essential oil had an efficacy of 78.6% against monogeneans. The toxicity of this essential oil was low, since there were a few physiological and histopathological changes that did not compromise the functioning of the gills of the fish. Therefore, 100 mg/L of P. hispidum essential oil was effective for controlling monogeneans in C. macropomum, when short therapeutic baths were used without compromising the health of the exposed fish. EO (EO spathulatus janauachensis macropomum fish 10 mgL mg L P 70 alcohol hday h day h/day 4 days 786 78 6 78.6 low Therefore C 7 78.

Resumo Este estudo investigou pela primeira vez, a eficácia de banhos terapêuticos com óleo essencial (OE) de Piper hispidum contra monogenéticos Anacanthorus spathulatus, Notozothecium janauachensis, Mymarothecium boegeri e Linguadactyloides brinkmanni das brânquias de Colossoma macropomum, bem como os efeitos hematológicos e histológicos neste peixe. Foram usados 100 mg/L de óleo essencial de P. hispidum e dois grupos controles (água do tanque de cultivo e água do tanque de cultivo com álcool 70%) expostos durante 1 hora por dia, com intervalos de 48 horas durante 3 dias, três repetições cada. Banhos terapêuticos com 100 mg/L de OE de P. hispidum tiveram uma eficácia de 78,6% contra monogenéticos. A toxicidade foi baixa, pois ocorreram poucas alterações fisiológicas e histopatológicas que não comprometeram o funcionamento das brânquias dos peixes. Portanto, houve eficácia de 100 mg/L de OE de P. hispidum para controle de monogenéticos em C. macropomum, quando usados banhos terapêuticos curtos sem prejudicar a saúde dos peixes expostos. vez (OE spathulatus janauachensis macropomum peixe 10 mgL mg L P 70% 70 dia 4 dias cada 786 78 6 78,6 baixa Portanto C 7 78,

ABSTRACT Synthetic chemical insecticides are widely used in population control of pests. Aedes aegypti is the primary vector of dengue, Zika, chikungunya and yellow fever to humans, and has proven resistance to chemical insecticides. As an alternative vector control method, the ethanolic extract of the leaves of Piper peltatum L. (Piperaceae) showed larvicidal activity against Ae. aegypti. Despite the wide medicinal use of this plant, the biological activity of its isolated constituents remains unexplored. In this sense, we isolated, identified and evaluated the larvicidal activity of 4-nerolidylcatechol (4-NC) from P. peltatum against Ae. aegypti, Culex quinquefasciatus and Anopheles darlingi, focusing on the larvicidal, adulticidal and genotoxic potential of 4-NC on Ae. aegypti. Larvae were captured in the city of Manaus, Amazonas state, Brazil. 4-NC was isolated from the extract of the leaves of P. peltatum via silica gel chromatography. This was identified using nuclear magnetic resonance spectroscopy and tested in Artemia franciscana (6.25, 12.5, 25, 50, 100 and 200 µg/mL). In the toxicity bioassay, Ae. aegypti larvae were exposed to 30, 50, 70, 90, and 110 µg/mL of 4-NC, while Cx. quinquefasciatus and An. darlingi were exposed to 6.25, 12.5, 25, 50 and 100 µg/mL. Ae. aegypti larvae were also subjected to 40 and 60 µg/mL of 4-NC (genotoxicity bioassay), and adult females to 62.5 to 1,000 µg/mL (adulticidal bioassay). The results of the 4-NC toxicity assays showed that there was 100% mortality in larvae of Ar. franciscana at the concentration of 200 µg/mL, with an LC50 of 8.0 μg/mL. In the larvae of Ae. aegypti, mortality was 100%, with an LC50 of 62 μg/mL. In larvae of Cx. quinquefasciatus, 97% mortality occurred, with an LC50 of 52.3 μg/mL, and in An. darlingi larvae there was an 83% mortality rate, with an LC50 of 55.8 μg/mL. In adults of Ae. aegypti, however, there was no adulticidal activity. In the larvae of Ae. aegypti, the genotoxic effect of 4-NC (40 and 60 µg/mL) showed significant frequency (p < 0.05) of cellular abnormalities (micronuclei, budding and nuclear bridges) of interphasic nuclei of neuroblasts and oocytes in relation to the negative control. This result may be associated with a decrease in oviposition of females, which was observed in two generations. We can confirm that 4-NC has larvicidal activity against Ae. aegypti, Cx. quinquefasciatus and An. darlingi. Although it does not present adulticidal activity in Ae. aegypti, it reduced the oviposition of females. Therefore, 4-NC seems to be a strong candidate for the development of an alternative method for the control of these mosquitoes in the immature phase. pests dengue Zika humans L Piperaceae (Piperaceae Ae plant unexplored sense 4nerolidylcatechol nerolidylcatechol 4 4NC NC (4-NC P Manaus state Brazil chromatography 625 6 25 (6.25 125 12 5 12.5 10 20 µgmL . µg mL bioassay 30 70 90 11 4NC, NC, Cx An 6.25 genotoxicity , bioassay) 62. 1000 1 000 1,00 bioassay. Ar LC LC5 80 8 0 8. μgmL μg μg/mL 97 occurred 523 52 3 52. 83 rate 558 55 55. however (4 p 0.05 005 05 micronuclei, micronuclei (micronuclei bridges generations Therefore phase 2 (6.2 12. 7 9 6.2 00 1,0 ( 0.0 (6. 6. 1, 0. (6

ABSTRACT Introduction: American trypanosomiasis or Chagas diseases, caused by Trypanosoma cruzi and Human African trypanosomiasis, caused by Trypanosoma brucei, are two neglected tropical, endemic and fatal disease, that affect 20 million people and cause 95,000 deaths per year in tropical and subtropical areas of the world. Objective: The objective of this review was to show the potential of the flora of Peru, emphasizing the Asteraceae family, against Trypanosoma cruzi, comparing it with American genera and species studied in various countries. Methods: A review of scientific articles on the subject was carried out, especially taken from the Scopus, SciELO, ScienceDirect, Medline and PubMed databases of American plant species, covering a period of twelve years (2010 to 2022). Results: A wide list of genera and species of the Asteraceae family and the flora of Peru with great potential in the treatment of trypanosomiasis is presented. Most of these species belong to the families Asteraceae, Piperaceae, Annonaceae, and Lauraceae. In the Asteraceae family, 48 genera and 76 species with trypanocidal activity were recorded, corresponding to 29 genera and 57 species for the flora of Peru. In these 29 genera, 174 endemic species potentially useful in the treatment of T. cruzi have been reported. Conclusions: The flora of Peru has around 22,000 species. Some of these genera and species are found in the flora of various American countries and have been tested in the treatment of Chagas disease, so the flora of Peru offers new opportunities in its treatment.

RESUMEN Introducción: La tripanosomiasis americana o enfermedad de Chagas, causada por la Trypanosoma cruzi y la tripanosomiasis africana humana, causada por la Trypanosoma brucei, son dos enfermedades tropicales desatendidas, endémicas y fatales, que afectan a 20 millones de personas y causan 95 000 muertes por año en áreas tropicales y subtropicales del mundo. Objetivos: El objetivo de esta revisión fue mostrar el potencial de la flora del Perú, enfatizar en la familia Asteraceae contra la Trypanosoma cruzi y compararlos con géneros y especies americanas estudiadas en diversos países. Métodos: Se realizó una revisión de artículos científicos sobre el tema de especies vegetales americanas, especialmente tomados de la base de datos Scopus, SciELO, ScienceDirect, Medline and PubMed; se abarcó un período de 12 años (2010 a 2022). Resultados: Se presenta una amplia relación de géneros y especies de la familia Asteraceae y de la flora del Perú con gran potencial en el tratamiento de la tripanosomiasis. La mayoría de estas especies pertenecen a las familias Asteraceae, Piperaceae, Annonaceae y Lauraceae. En la familia Asteraceae se registraron 48 géneros y 76 especies con actividad tripanocida; correspondieron 29 géneros y 57 especies para la flora del Perú. Asimismo, en estos 29 géneros se han reportado 174 especies endémicas potencialmente útiles en el tratamiento de la T. cruzi. Conclusiones: La flora del Perú tiene alrededor de 22 000 especies. Algunos de estos géneros y especies se encuentran en la flora de varios países americanos y han sido ensayados en el tratamiento de la enfermedad de Chagas, por lo que la flora del Perú ofrece nuevas oportunidades en su tratamiento.

RESUMEN Los bosques montanos se encuentran en un rango altitudinal entre 800 y 3500 m snm, conservan una importante diversidad de especies arbóreas y altos niveles de endemismos. Muchos de estos bosques están siendo convertidos a otros usos de tierra y en algunos casos ocurren procesos sucesionales que permiten el establecimiento de bosques secundarios. Con el fin de evaluar la contribución florística de estos bosques montanos secundarios a la flora arbórea de la región de la Selva Central del Perú, se analizó la composición florística y la diversidad arbórea en dos bosques secundarios ubicados en la provincia de Chanchamayo. Para ello, se instalaron dos parcelas per- manentes de monitoreo de vegetación, siendo estas la Parcela Puyu Sacha Bosque Secundario (P-PS) de 1 ha ubicada en un bosque montano a 2060 m snm, donde se registraron 638 individuos, 52 especies, 37 géneros y 28 familias botánicas, y la Parce- la Génova Bosque Secundario 2 (P-GS2) de 0,6 ha ubicada en un bosque premontano a 1040 m snm, donde se registraron 242 individuos, 34 especies, 32 géneros y 21 familias botánicas. En P-PS (estrato montano entre 1500 y 3500 m snm) las familias más abundantes fueron Urticaceae, Actinidiaceae, Melastomataceae, Cordiaceae y Rubiaceae; y las especies más abundantes fueron Cordia cylindristachya (Ruiz & Pav.) Roem. & Schult., Miconia aff. crassistigma Cogn., Saurauia biserrata (Ruiz & Pav.) Spreng., Cecropia angustifolia Trécul, Cecropia sp.2 y Saurauia glabra (Ruiz & Pav.) Soejarto. En P-GS2 (estrato premontano entre 800 y 1500 m snm) las familias más abundantes fueron Euphorbiaceae, Fabaceae, Cannabaceae, Piperaceae y Urticaceae; y las especies más abundantes fueron Sapium glandulosum (L.) Morong, Trema mi- crantha (L.) Blume, Cecropia polystachya Trécul, Erythrina ulei Harms y Piper aduncum L. Ambas parcelas presentaron bajos niveles de diversidad debido a que se ubican sobre bosques secundarios que en el pasado tuvieron intervenciones antrópicas.

ABSTRACT Montane forests are found in an altitudinal range between 800 and 3500 m asl and conserve an important diversity of tree species and high levels of endemism. Many of these forests are being converted to other land uses and, in some cases, successional processes are occurring that allow the establishment of secondary forests. In order to evaluate the floristic contribution of these secondary montane forests to the arboreal flora of the Selva Central region of Peru, we analyzed the floristic composition and tree diversity in two secondary forests located in the province of Chanchamayo. For this purpose, two permanent vegetation monitoring plots were installed, the Puyu Sacha Secondary Forest Plot (P-PS) of 1 ha located in a montane forest at 2060 m asl, where 638 individuals, 52 species, 37 genera and 28 botanical families were recorded, and the Genova Secondary Forest 2 Plot (P-GS2) of 0.6 ha located in a premontane forest at 1040 m asl, where 242 individuals, 34 species, 32 genera and 21 botanical families were recorded. In P-PS (montane stratum between 1500 and 3500 m asl) the most abundant families were Urticaceae, Actinidiaceae, Melastomataceae, Cordiace- ae and Rubiaceae; and the most abundant species were Cordia cylindristachya (Ruiz & Pav.) Roem. & Schult., Miconia aff. crassistigma Cogn., Saurauia biserrata (Ruiz & Pav.) Spreng., Cecropia angustifolia Trécul, Cecropia sp.2 and Saurauia glabra (Ruiz & Pav.) Soejarto. In P-GS2 (premontane stratum between 800 and 1500 m asl) the most abundant families were Euphorbiaceae, Fabaceae, Cannabaceae, Piperaceae and Urticaceae; and the most abundant species were Sapium glandulosum (L.) Morong, Trema micrantha (L.) Blume, Cecropia polystachya Trécul, Erythrina ulei Harms and Piper aduncum L. Both plots showed low levels of diversity because they are located in secondary forests that in the past had anthropic interventions.

RESUMO O objetivo desta pesquisa foi investigar a influência de técnicas de secagem de folhas (bancada e em estufa a 35 e 45 °C) sobre o rendimento do óleo essencial (OE) de Piper aduncum L., Piper anonifolium Kunth, Piper crassinervium Kunth e Piper hispidinervum C. DC.; analisar o perfil químico dos OEs; e o potencial inseticida destes óleos para Ascia monuste orseis (Godart), Atta sexdens L., e para Zabrotes subfasciatus (Boheman), para o besouro Cryptolestes ferrugineus (Stephens) e para Sitophilus zeamais Motschulsky. Avaliou-se o rendimento do OE utilizando quatro repetições de 100g de folhas secas. Os OEs foram obtidos por hidrodestilação e submetidos à análise por CG-EM para a constatação da composição química. Utilizou-se as concentrações 2,60 e 157,25 nL/cm2 nos bioensaios de toxicidade dos óleos. O rendimento dos OEs foi maior nas espécies P. aduncum e P. hispidinervum utilizando folhas secas em estufa a 45 °C, com rendimentos médios de 4,72±0,04% e 2,61±0,26% respectivamente. Os constituintes majoritários presentes nos OEs de P. hispidinervum e P. aduncum, foram o Safrol (98,80%) e Apiole (90,00%). Para P. anonifolium, foram o α-Muuroleno (23,11%), y-Muuroleno (16,60%) e Cadina-1(10), enquanto para P. crassinervium, foram o Viridiflorol (27,70%) e Sabineno (15,50%). Constatou-se que os OEs de P. aduncum, P. anonifolium, P. crassinervium e P. hispidinervum apresentaram efeito tóxico para os insetos, exceto P. anonifolium e P. crassinervium para S. zeamais. O rendimento dos OEs foi maior nas espécies P. aduncum e P. hispidinervum e estes óleos causaram maior taxa de mortalidade para os insetos investigados. bancada 3 4 °C C (OE L L. DC. DC Godart, Godart , (Godart) Boheman, Boheman (Boheman) Stephens (Stephens Motschulsky Avaliouse Avaliou se g CGEM CG EM química Utilizouse Utilizou 260 2 60 2,6 15725 157 25 157,2 nLcm2 nLcm nL cm2 cm nL/cm P 472004 72 0 04 4,72±0,04 261026 61 26 2,61±0,26 respectivamente 98,80% 9880 98 80 (98,80% 90,00%. 9000 90,00% . 90 00 (90,00%) αMuuroleno α Muuroleno 23,11%, 2311 23,11% 23 11 (23,11%) yMuuroleno y 16,60% 1660 16 (16,60% Cadina110, Cadina110 Cadina 1 10 Cadina-1(10) 27,70% 2770 27 70 (27,70% 15,50%. 1550 15,50% 15 50 (15,50%) Constatouse Constatou S investigados (Godart (Boheman 6 2, 1572 157, 47200 7 4,72±0,0 26102 2,61±0,2 98,80 988 9 8 (98,80 900 90,00 (90,00% 231 23,11 (23,11% 16,60 166 (16,60 Cadina11 Cadina-1(10 27,70 277 (27,70 155 15,50 5 (15,50% 4720 4,72±0, 2610 2,61±0, 98,8 (98,8 90,0 (90,00 23,1 (23,11 16,6 (16,6 Cadina1 Cadina-1(1 27,7 (27,7 15,5 (15,50 472 4,72±0 261 2,61±0 98, (98, 90, (90,0 23, (23,1 16, (16, Cadina-1( 27, (27, 15, (15,5 47 4,72± 2,61± (98 (90, (23, (16 Cadina-1 (27 (15, 4,72 2,61 (9 (90 (23 (1 Cadina- (2 (15 4,7 ( 4,

ABSTRACT The objective of this study was to investigate the influence of leaf drying techniques (bench and oven at 35 and 45 °C) on the essential oil (EO) yield of Piper aduncum L., Piper anonifolium Kunth, Piper crassinervium Kunth and Piper hispidinervum C. DC., and to analyze the chemical profile of EOs and the insecticidal potential of these oils against Ascia monuste orseis (Godart), Atta sexdens L., Zabrotes subfasciatus (Boheman), Cryptolestes ferrugineus (Stephens) and Sitophilus zeamais Motschulsky. EO yield was evaluated using four replicates of 100g of dry leaves. The EOs were obtained by hydrodistillation and subjected to GC-MS analysis to assess the chemical composition. Concentrations of 2.60 and 157.25 nL/cm2 were used in the oil toxicity bioassays. EO yield was higher in the species P. aduncum and P. hispidinervum using leaves dried in oven at 45 °C, with average yields of 4.72±0.04% and 2.61±0.26%, respectively. The major constituents present in the EOs of P. hispidinervum and P. aduncum were Safrole (98.80%) and Apiole (90.00%). For P. anonifolium, the major constituents were α-Muurolene (23.11%), y-Muurolene (16.60%) and Cadina-1(10), while for P. crassinervium, they were Viridiflorol (27.70%) and Sabinene (15.50%). It was found that the EOs of P. aduncum, P. anonifolium, P. crassinervium and P. hispidinervum had a toxic effect on insects, except for P. anonifolium and P. crassinervium for S. zeamais. EO yield was higher in the species P. aduncum and P. hispidinervum, and these oils caused a higher mortality rate for the investigated insects. bench 3 4 °C C (EO L L. DC DC. Godart, Godart , (Godart) Boheman, Boheman (Boheman) Stephens (Stephens Motschulsky g GCMS GC MS composition 260 2 60 2.6 15725 157 25 157.2 nLcm2 nLcm nL cm2 cm nL/cm bioassays P 472004 72 0 04 4.72±0.04 261026 61 26 2.61±0.26% respectively 98.80% 9880 98 80 (98.80% 90.00%. 9000 90.00% . 90 00 (90.00%) αMuurolene α Muurolene 23.11%, 2311 23.11% 23 11 (23.11%) yMuurolene y 16.60% 1660 16 (16.60% Cadina110, Cadina110 Cadina 1 10 Cadina-1(10) 27.70% 2770 27 70 (27.70% 15.50%. 1550 15.50% 15 50 (15.50%) insects S (Godart (Boheman 6 2. 1572 157. 47200 7 4.72±0.0 26102 2.61±0.26 98.80 988 9 8 (98.80 900 90.00 (90.00% 231 23.11 (23.11% 16.60 166 (16.60 Cadina11 Cadina-1(10 27.70 277 (27.70 155 15.50 5 (15.50% 4720 4.72±0. 2610 2.61±0.2 98.8 (98.8 90.0 (90.00 23.1 (23.11 16.6 (16.6 Cadina1 Cadina-1(1 27.7 (27.7 15.5 (15.50 472 4.72±0 261 2.61±0. 98. (98. 90. (90.0 23. (23.1 16. (16. Cadina-1( 27. (27. 15. (15.5 47 4.72± 2.61±0 (98 (90. (23. (16 Cadina-1 (27 (15. 4.72 2.61± (9 (90 (23 (1 Cadina- (2 (15 4.7 2.61 ( 4.

Resumo Piper aduncum L., uma planta medicinal brasileira, é conhecida por suas propriedades bioativas, incluindo efeitos repelentes e inseticidas. Este estudo investigou o potencial inseticida dos óleos essenciais (OEs) de P. aduncum, coletados na estação seca e chuvosa, contra pulgas (Ctenocephalides felis felis Bouché, 1835) em estágios de ovos e adultos. Os OEs foram obtidos por hidrodestilação em aparelho de Clevenger, modificado por 2 horas. Análises qualitativas e quantitativas foram realizadas por meio de cromatografia gasosa. Os resultados revelaram que o dilapiol foi a substância predominante, representando 77,56% (chuvoso) e 85,52% (seco) dos OEs. Esses OEs mostraram alta eficácia contra o parasita C. felis felis, resultando em 100% de mortalidade de ovos a uma concentração de 100 μg/mL e 100% de mortalidade para pulgas adultas a partir de 1.000 μg/mL. O padrão dilapiol também foi eficaz, mas em uma concentração relativamente alta. Este achado sugere que os OEs de P. aduncum exibem citotoxicidade contra esses parasitas e podem ter potencial para a produção comercial, oferecendo aplicações práticas para essa bioprospecção. Esse estudo revelou de maneira única que os OEs de P. aduncum, ricos em dilapiol, demonstraram atividade pulicida contra o parasita C. felis felis, inibindo, especialmente, a eclosão dos ovos desses parasitas. L L. brasileira bioativas inseticidas (OEs P chuvosa Ctenocephalides Bouché 1835 adultos Clevenger horas gasosa predominante 7756 77 56 77,56 chuvoso (chuvoso 8552 85 52 85,52 seco (seco C 10 μgmL μg mL 1000 1 000 1.00 eficaz comercial bioprospecção inibindo especialmente 183 775 7 5 77,5 855 8 85,5 00 1.0 18 77, 85, 0 1.

Abstract Piper aduncum L., a Brazilian medicinal plant, is known for its bioactive properties, including repellent and insecticidal effects. This study investigated the insecticidal potential of essential oils (EOs) from P. aduncum, collected during the dry and rainy seasons, against fleas (Ctenocephalides felis felis Bouché, 1835) in egg and adult stages. The EOs were obtained by hydrodistillation using a modified Clevenger apparatus for 2 h. Qualitative and quantitative analysis were performed via gas chromatography. The findings revealed that dillapiole was the predominant substance in both EOs, accounting for 77.6% (rainy) and 85.5% (dry) of the EOs. These EOs exhibited high efficacy against the parasite C. felis felis, resulting in 100% egg mortality at a concentration of 100 μg/mL and 100% mortality for adult fleas starting from 1,000 μg/mL. Dillapiole standard was also effective but at a relatively high concentration. This finding suggested that EOs from P. aduncum exhibit cytotoxicity against these pests and might hold potential for commercial production, offering practical applications for such bioprospecting. This study uniquely revealed that the EOs from P. aduncum, which is rich in dillapiole, demonstrated pulicidal activity against the parasite C. felis felis, particularly in inhibiting the hatching of the eggs of these parasites. L L. plant properties effects (EOs P seasons Ctenocephalides Bouché 1835 stages h chromatography 776 77 6 77.6 (rainy 855 85 5 85.5 (dry C 10 μgmL μg mL 1000 1 000 1,00 production bioprospecting parasites 183 7 77. 8 85. 00 1,0 18 0 1,

Abstract Piper species are important components of the understory in tropical and subtropical forests, and some species are of economic and medicinal importance. This genus has been studied in different areas of science. However, some questions remain unanswered after the new circumscription as proposed by phylogenetic studies. Here, I review different aspects related to the reproductive biology of Piper species (phenology, sexual expression, floral biology, pollination and mating systems), comparing the results between Neotropical (representing most of the studies carried out so far) and Paleotropical regions. Seventy-five species were analyzed. Some patterns can be observed, mainly in Neotropical species, such as the predominance of annual flowering and fruiting, dichogamy (generally incomplete protogyny) with gradual and sequential exposure of stigmas, asynchronous pollen release, and pollination by insects. Some aspects have been poorly studied, such as variations in sexual expression, the efficiency of different pollinators, and the chemical composition of floral volatiles associated with their role in pollination. In addition, I discuss remaining gaps and further studies that are required, mainly on Paleotropical Piper species, to obtain basic information on the reproductive biology of these plants. From this, comparisons with Neotropical species can be made, allowing the verification of patterns in this genus. forests importance science However Here phenology, phenology (phenology expression systems, systems , systems) representing far regions Seventyfive Seventy five analyzed observed fruiting generally protogyny stigmas release insects pollinators addition required plants made

Resumo Espécies de Piper são importantes componentes do sub-bosque em florestas tropicais e subtropicais, e algumas espécies são de importância econômica e medicinal. Esse gênero tem sido estudado em diferentes áreas da ciência. No entanto, algumas questões permanecem sem resposta após a nova circunscrição proposta pelos estudos filogenéticos. Aqui, reviso diferentes aspectos relacionados à biologia reprodutiva das espécies de Piper (fenologia, expressão sexual, biologia floral, sistemas de polinização e reprodutivo), comparando os resultados entre as regiões neotropical (representando a maioria dos estudos realizados até agora) e paleotropical. 75 espécies foram analisadas. Alguns padrões podem ser observados, principalmente em espécies neotropicais, como a predominância de floração e frutificação anual, dicogamia (geralmente protoginia incompleta) com exposição gradual e sequencial dos estigmas, liberação assíncrona de pólen e polinização por insetos. Alguns aspectos têm sido pouco estudados, como as variações na expressão sexual, a eficiência dos diferentes polinizadores e a composição química dos voláteis florais associados ao seu papel na polinização. Além disso, discuto as lacunas remanescentes e são necessários mais estudos, principalmente sobre as espécies de Piper paleotropicais, para obter informações básicas sobre a biologia reprodutiva dessas plantas. A partir disso, comparações com espécies neotropicais podem ser feitas, permitindo a verificação de padrões nesse gênero. subbosque sub bosque subtropicais medicinal ciência entanto filogenéticos Aqui fenologia, fenologia (fenologia sexual floral reprodutivo, reprodutivo , reprodutivo) representando agora paleotropical 7 analisadas observados anual geralmente incompleta estigmas insetos estudados disso paleotropicais plantas feitas

ABSTRACT Purpose: To assess the effect of the ethanolic extract of the leaves of three species of plants from the Piperaceae family on reducing necrosis and enhancing wound healing in an animal model of degloving injuries. Methods: The animals were divided into six groups, each consisting of six animals: sham, negative control, EEPA (Piper amalago ethanolic extract), EEPG (Piper glabratum ethanolic extract), EEPV (Piper vicosanum ethanolic extract), and positive control receiving hyperbaric oxygenation. The animals underwent surgery to induce excision wounds, and the extent of cutaneous necrosis was evaluated using graphic software, while wound healing was assessed through histopathology. Results: Skin necrosis percentage area was: sham group = 62.84% 6.38; negative control group = 63.03% 4.11; P. vicosanum = 40.80% 4.76 p < 0.05; P. glabratum 32.97% 4.01 p < 0.01; P. amalago = 32.40% 4.61 p < 0.01; hyperbaric oxygenation = 33.21% 4.29 p < 0.01. All treated groups showed higher collagen deposition and less intense, plus predominantly mononuclear inflammatory infiltrate, suggesting improved healing process. Conclusions: The three tested extracts demonstrated efficacy in reducing the extent of cutaneous necrosis caused by degloving injuries and also showed evidence of improvement in the wound healing process. Purpose Methods Piper extract, , extract) wounds software histopathology Results 6284 62 84 62.84 6.38 638 6 38 6303 63 03 63.03 4.11 411 4 11 P 4080 40 80 40.80 476 76 4.7 0.05 005 0 05 3297 32 97 32.97 401 01 4.0 0.01 001 3240 32.40 461 61 4.6 3321 33 21 33.21 429 29 4.2 intense infiltrate process Conclusions 628 8 62.8 6.3 3 630 63.0 4.1 41 1 408 40.8 47 7 4. 0.0 00 329 9 32.9 324 32.4 46 332 2 33.2 42 62. 6. 63. 40. 0. 32. 33.

ABSTRACT Determining the bioactivity of essential oils extracted from plants is crucial for the development of alternatives for controlling insect pests in stored grains. This study aimed to investigate the bioactivity of Piper aduncum L. essential oil for the control of Sitophilus zeamais (Coleoptera: Curculionidae), Sitophilus oryzae (Coleoptera: Curculionidae) and Cryptolestes ferrugineus (Coleoptera: Cucujidae). Bioassays were conducted to determine the oil toxicity, sublethal effects, attractiveness and flight initiation behavior. Toxicity tests were carried out to determine the lethal concentrations for 50 and 95 % of the insects (LC50 and LC95). Tests were also performed with sublethal exposures to C. ferrurineus, S. oryzae and S. zeamais, using their respective LC5 values (0.0002, 0.097 and 0.11 µL cm-2), to investigate effects on the attraction or repellency activity, as well as flight tests. C. ferrugineus showed greater LC50 (0.0005 µL cm-2) and LC95 (0.0012 µL cm-2) susceptibility, in relation to S. oryzae LC50 (0.35 µL cm-2) and LC95 (1.26 µL cm-2), and S. zeamais LC50 (0.39 µL cm-2) and LC95 (1.40 µL cm-2). The exposure to the sublethal LC5 concentration influenced the behavioral responses of attractiveness/repellency and flight activity. The oil had a neutral effect on C. ferrugineus LC5 (0.0002 µL cm-2) and an attractive effect on S. oryzae and S. zeamais LC5 (0.097 and 0.11 µL cm-2), and can be considered a potential insecticide for controlling pest weevil species of stored grains. grains L Coleoptera (Coleoptera Curculionidae, Curculionidae , Cucujidae. Cucujidae . Cucujidae) toxicity behavior 5 9 LC (LC5 LC95. LC95) C ferrurineus S 0.0002, 00002 0 0002 0097 097 0.09 011 11 0.1 cm2, cm2 cm 2 activity 0.0005 00005 0005 (0.000 cm-2 LC9 0.0012 00012 0012 (0.001 susceptibility 0.35 035 35 (0.3 1.26 126 1 26 (1.2 0.39 039 39 1.40 140 40 (1.4 cm2. attractivenessrepellency 0.0002 (0.09 (LC 0000 000 009 09 0.0 01 0. 0.000 (0.00 cm- 0.001 0001 001 0.3 03 3 (0. 1.2 12 (1. 1.4 14 4 (0.0 00 0.00 (0 1. (1 (

RESUMO Determinar a bioatividade de óleos essenciais extraídos de plantas é primordial para o desenvolvimento de alternativas de controle de insetos-praga em grãos armazenados. Objetivou-se investigar a bioatividade do óleo essencial de Piper aduncum L. para o controle de Sitophilus zeamais (Coleoptera: Curculionidae), Sitophilus oryzae (Coleoptera: Curculionidae) e Cryptolestes ferrugineus (Coleoptera: Cucujidae). Foram realizados bioensaios para determinar a toxicidade do óleo, efeitos subletais, comportamento de atratividade e iniciação de voo. Testes de toxicidade foram realizados, determinando-se as concentrações letais para 50 e 95 % dos insetos (CL50 e CL95). Também foram realizados testes com exposições subletais para C. ferrurineus, S. oryzae e S. zeamais, utilizando-se as suas respectivas CL5 (0,0002; 0,097; e 0,11 µL cm-2), para averiguar efeitos na atividade de atração ou repelência, e testes de voo. C. ferrugineus apresentou maior susceptibilidade CL50 (0,0005 µL cm-2) e CL95 (0,0012 µL cm-2), em relação a S. oryzae CL50 (0,35 µL cm-2) e CL95 (1,26 µL cm-2) e S. zeamais CL50 (0,39 µL cm-2) e CL95 (1,40 µL cm-2). A exposição à concentração subletal CL5 influenciou nas respostas comportamentais de atratividade/repelência e atividade de voo das espécies. O óleo apresentou efeito neutro para C. ferrugineus CL5 (0,0002 µL cm-2) e atrativo para S. oryzae e S. zeamais CL5 (0,097 e 0,11 µL cm-2), podendo ser considerado potencial inseticida para o controle de espécies de gorgulhos-praga de grãos armazenados. insetospraga praga armazenados Objetivouse Objetivou se L Coleoptera (Coleoptera Curculionidae, Curculionidae , Cucujidae. Cucujidae . Cucujidae) determinandose determinando 5 9 CL (CL5 CL95. CL95) C ferrurineus S utilizandose utilizando 0,0002 00002 0 0002 0,097 0097 097 011 11 0,1 cm2, cm2 cm 2 repelência 0,0005 00005 0005 (0,000 cm-2 CL9 0,0012 00012 0012 (0,001 0,35 035 35 (0,3 1,26 126 1 26 (1,2 0,39 039 39 1,40 140 40 (1,4 cm2. atratividaderepelência (0,09 gorgulhospraga gorgulhos (CL 0,000 0000 000 0,09 009 09 01 0, (0,00 cm- 0,001 0001 001 0,3 03 3 (0, 1,2 12 (1, 1,4 14 4 (0,0 0,00 00 0,0 (0 1, (1 (

Abstract This literature brief review compiled the main biological activities and active chemical constituents that have already been isolated from Piper aduncum (Piperaceae), whose common name is spiked paper. In Brazilian Portuguese, it has been called pimenta-longa, aperta-ruão, pimenta de macaco, matico, erva de jaboti, jaborandi do mato, pimenta-de-fruto-ganchoso and tapa buraco. Extracts, essential oils (EOs) and compounds isolated from this species have exhibited remarkable fungicidal and insecticidal activities, besides antibacterial, antileishmanial, antioxidant, cytotoxic/antitumor, larvicidal, antiplatelet, molluscoidal and antiviral ones. P. aduncum is an endemic species in Brazil which has drawn researchers’ attention because of high contents of EOs that are extracted from its leaves, inflorescences and twigs. This study aimed at analyzing data on P. aduncum, its EOs, extracts and isolated compounds found in 45 research papers. Its major chemical constituent is definitely dillapiole, an arylpropanoid. Findings were carefully compiled by this brief review which reinforces the chemical and biological potential of this botanic species. Piperaceae, Piperaceae , (Piperaceae) paper Portuguese pimentalonga, pimentalonga longa, longa pimenta-longa apertaruão, apertaruão aperta ruão, ruão aperta-ruão macaco matico jaboti mato pimentadefrutoganchoso fruto ganchoso buraco Extracts (EOs antibacterial antileishmanial antioxidant cytotoxicantitumor cytotoxic antitumor cytotoxic/antitumor larvicidal antiplatelet ones P researchers leaves twigs 4 papers dillapiole arylpropanoid (Piperaceae

RESUMEN Los ecosistemas andinos constituyen uno de los más importantes centros de biodiversidad del mundo, sin embargo, sus coberturas boscosas tienden a desaparecer rápidamente debido a que están sometidas a una fuerte presión antrópica. En este estudio, se caracterizan las angiospermas presentes en los parches de bosque, sus bordes y en zonas de vegetación abierta o potrero de un área andina de la Cordillera Central de Colombia. Entre marzo de 2017 y febrero de 2018 se realizaron recolecciones de material fértil mediante recorridos del área. Se obtuvo información complementaria de colecciones históricas depositadas en herbarios. El material herborizado se depositó en el Herbario Nacional Colombiano (COL). Se registran 431 especies, agrupadas en 92 familias y 239 géneros, se encontraron cinco familias de magnólidas con siete géneros y 29 especies, 18 de monocotiledóneas (66/131) y 69 eudicotiledóneas (166/271). La mayor riqueza a nivel de especies la presentan Orchidaceae (54), Rubiaceae (24), Asteraceae (22), Araceae (22) y Piperaceae (20). Las hierbas terrestres son la forma de vida más numerosa (31,8 %), seguidas por arbustos (26,5 %) y árboles (17,6 %). A pesar de tratarse de áreas rodeadas por actividades productivas, principalmente agropecuarias, se encontraron 30 especies endémicas de Colombia, 170 taxones representan nuevos registros para la vertiente oriental de la Cordillera Central del departamento de Antioquia, dos nuevos registros para el departamento y cuatro nuevas especies.

ABSTRACT The Andean ecosystems constitute one of the most important centers of biodiversity in the world; however, their forest covers tend to disappear rapidly as they are subjected to strong anthropic pressure. In this study, the angiosperms present in the forest patches, its edge, and in areas of open vegetation or pasture of an Andean area of the Central Cordillera of Colombia are characterized. General collections of fertile material were made through tours between March 2017 and February 2018. Complementary information was obtained from historic collections deposited in herbaria and the botanical material was deposited in the Herbario Nacional Colombiano (COL). A total of 431 species are recorded, grouped into 92 families and 239 genera, five families of magnolids were found with seven genera and 29 species, 18 monocotyledons (66/131) and 69 eudicotyledons (166/271). The highest species richness is presented by Orchidaceae (54), Rubiaceae (24), Asteraceae (22), Araceae (22), and Piperaceae (20). Terrestrial herbs is the most numerous life form (31.8 %), followed by shrubs (26.5 %) and trees (17.6 %). Despite being areas surrounded by productive activities, mainly agricultural, 30 species are endemic to Colombia, 170 taxa are new records for the eastern slope of the Central mountain range of the department of Antioquia, two are new records for the department, and four new species are reported.

Abstract Several Piper species accumulate piperamides as secondary metabolites, and although they have relevant biological importance, many details of their biosynthetic pathways have not yet been described experimentally. Experiments involving enzymatic reactions and labeled precursor feeding were performed using the species Piper tuberculatum and Piper arboreum. The activities of the phenylalanine ammonia lyase (PAL) enzymes, which are involved in the general phenylpropanoid pathway, were monitored by the conversion of the amino acid L-phenylalanine to cinnamic acid. The activity of the 4-hydroxylase (C4H) enzyme was also observed in P. tuberculatum by converting cinnamic acid to p-coumaric acid. L-[UL-14C]-phenylalanine was fed into the leaves of P. tuberculatum and incorporated into piperine (1), 4,5-dihydropiperine (2), fagaramide (4), trans-piplartine (7), and dihydropiplartine (9). In P. arboreum, it was only incorporated into the piperamide 4,5-dihydropiperiline (3). L-[UL-14C]-lysine was successfully incorporated into the 4,5-dihydropiperine piperidine group (2), dihydropyridinone, and trans- (7) and cis-piplartine (8). These data corroborate the proposal of mixed biosynthetic origin of piperamides with the aromatic moiety originating from cinnamic acid (shikimic acid pathway) and key amide construction with amino acids as precursors. metabolites importance experimentally arboreum PAL (PAL enzymes pathway Lphenylalanine L 4hydroxylase hydroxylase 4 C4H CH C H (C4H P pcoumaric p coumaric LUL14Cphenylalanine LULCphenylalanine UL 14C 1, 1 , (1) 4,5dihydropiperine 45dihydropiperine dihydropiperine 4,5 5 2, 2 (2) 4, (4) transpiplartine trans piplartine 7, 7 9. 9 . (9) 4,5dihydropiperiline 45dihydropiperiline dihydropiperiline 3. 3 (3) LUL14Clysine LULClysine lysine dihydropyridinone (7 cispiplartine cis 8. 8 (8) shikimic precursors LUL Cphenylalanine (1 5dihydropiperine 45 (2 (4 (9 5dihydropiperiline (3 Clysine ( (8

Resumo Diversas espécies do gênero Piper acumulam piperamidas como metabólitos secundários e, embora tenham relevante importância biológica, detalhes de suas vias biossintéticas ainda não foram descritos experimentalmente. Experimentos envolvendo reações enzimáticas e incorporação de precursores marcados foram realizados utilizando as espécies Piper tuberculatum e Piper arboreum. As atividades das enzimas fenilalanina amônia liase (PAL), que estão envolvidas na via geral dos fenilpropanoides, foram monitoradas pela conversão do aminoácido L-fenilalanina em ácido cinâmico. A atividade da enzima 4-hidroxilase (C4H) também foi observada em P. tuberculatum pela conversão do ácido cinâmico em ácido p-cumárico. L-[UL-14C]-fenilalanina foi administrada em folhas de P. tuberculatum e incorporada em piperina (1), 4,5-di-hidropiperina (2), fagaramida (4), trans-piplartina (7) e diidropiplartina (9). Em P. arboreum, foi incorporado apenas à piperamida 4,5-dihidropiperilina (3). L-[UL-14C]-lisina foi incorporada com sucesso em 4,5-diidropiperina (2), trans-piplartina (7) e cis-piplartina (8). Esses dados corroboram a proposta de origem biossintética mista de piperamidas com a porção aromática originada do ácido cinâmico (via do ácido chiquímico) e construção de amida chave com aminoácidos como precursores. biológica experimentalmente arboreum PAL, PAL , (PAL) fenilpropanoides Lfenilalanina L 4hidroxilase hidroxilase 4 C4H CH C H (C4H P pcumárico. pcumárico p cumárico. cumárico p-cumárico LUL14Cfenilalanina LULCfenilalanina UL 14C 1, 1 (1) 4,5dihidropiperina 45dihidropiperina dihidropiperina 4,5 di hidropiperina 5 2, 2 (2) 4, (4) transpiplartina trans piplartina 7 (7 9. 9 . (9) 4,5dihidropiperilina 45dihidropiperilina dihidropiperilina 3. 3 (3) LUL14Clisina LULClisina lisina 4,5diidropiperina 45diidropiperina diidropiperina cispiplartina cis 8. 8 (8) chiquímico (PAL LUL Cfenilalanina (1 5dihidropiperina 45 (2 (4 ( (9 5dihidropiperilina (3 Clisina 5diidropiperina (8