Inheritance of seedlessness and the molecular characterization of the INO gene in Annonaceae / Herança da ausência de sementes e a caracterização molecular do gene INO em Annonaceae

Abstract The inheritance of the seedless fruit characteristic of Annona squamosa has not yet been explained. Molecular techniques may aid breeding programs, mainly in the assisted selection of the target gene. The INO gene may be related to seed development in these fruits. The objective of the present paper was to investigate the inheritance of seedlessness in the 'Brazilian seedless' sugar apple and INO gene conservation in Annona squamosa and Annona cherimola x Annona squamosa genotypes by assessing their homology with the INO database genes. The F1 generation was obtained by crossing the mutant 'Brazilian seedless' (male genitor) (P1) with the wild-type A. squamosa with seeds (M1 and M2, female genitors). The INO gene was studied in mutant and wild-type A. squamosa (P1, M1, M2 and M3) and in the Gefner atemoya (A. cherimola x A. squamosa) (M4) cultivar. The DNA was extracted from young leaves, and four sets of specific primers flanking the INO gene were amplified. The seedless characteristic was identified as stenospermatic in the fruits of parental P1, suggesting monogenic inheritance with complete dominance. High sequence similarity of the INO gene amplifications in the sugar apple accessions (M1, M2, M3) and the atemoya cultivar Gefner (M4) reinforces the hypothesis of their conservation.

Resumo A herança da característica de fruto sem sementes de Annona squamosa ainda não foi esclarecida. Técnicas moleculares podem auxiliar em programas de melhoramento, principalmente na seleção assistida do gene de interesse. O gene INO pode estar relacionado ao desenvolvimento da semente dessas frutas. O objetivo foi investigar a herança da ausência de sementes em Annona squamosa e a conservação do gene INO nos genótipos Annona squamosa e Annona cherimola x Annona squamosa avaliando sua homologia com banco de dados de genes INO. A geração F1 foi obtida pelo cruzamento do mutante 'Brazilian seedless' (genitor masculino) (P1) com o tipo selvagem com sementes (A. squamosa) (M1 e M2, genitores femininos). O gene INO foi estudado em A. squamosa, mutante e selvagem (P1, M1, M2 e M3) e na cultivar Gefner atemoya (A. cherimola x A. squamosa) (M4). O DNA foi extraído de folhas jovens, e quatro conjuntos de primers específicos flanqueando o gene INO foram amplificados. A característica sem sementes foi identificada como estenospermática nos frutos do parental P1, sugerindo herança monogênica com dominância completa. A alta similaridade de sequência das amplificações do gene INO nos acessos de pinha (M1, M2, M3) e na cultivar de atemóia Gefner (M4) reforça a hipótese de sua conservação.

Uso tradicional de plantas medicinales por el adulto mayor en la comunidad Lechuga. Arroyo Naranjo / Traditional use of medicinal plants for the major adult in the Lechuga community. Arroyo Naranjo

Introducción: el uso de las plantas medicinales es común en la población de Cuba. Pero, gran parte de la misma desconoce sus propiedades, forma de empleo, modo de aplicación, así como las contraindicaciones e interacciones con los medicamentos convencionales. En este contexto el adulto mayor juega un rol importante por los conocimientos empíricos y esotéricos adquiridos, además del empleo que hace de estas plantas. Objetivo: valorar el uso tradicional de plantas medicinales por el adulto mayor en la comunidad Lechuga de Arroyo Naranjo. Métodos: entrevista, basada en una guía de aspectos y temáticas específicas a adultos mayores, de la comunidad Lechuga de Arroyo Naranjo. Se obtuvo una lista de plantas medicinales con frecuente uso, sus aplicaciones, partes utilizadas, modos de empleo y conocimientos sobre sus contraindicaciones, toxicidad o interacciones con medicamentos convencionales. Resultados: se alegó el uso de 38 especies de plantas medicinales, agrupadas en 25 familias, el 71 % de las plantas informadas se cultivaban en los alrededores de las casas, el 65,9 % de las personas conoce un número importante de las propiedades y usos de las plantas medicinales, el 56,8 % expuso al menos una forma o tipo de preparación de las plantas identificadas. Se constató que las mujeres conocen más de las propiedades y usos de las plantas medicinales que los hombres, el 85,2 % no conoce las contraindicaciones, toxicidad, riesgos, así como las posibles interacciones con los medicamentos convencionales. Conclusiones: el adulto mayor en la comunidad de Lechuga, tiene conocimientos empíricos sobre el uso de plantas con propiedades medicinales, aunque es necesario la divulgación y educación de la población en relación con las propiedades útiles de las plantas, formas de preparación y contraindicaciones, toxicidad o interacciones con medicamentos convencionales.

Interaction of Bephratelloides cubensis (Hymenoptera: Eurytomidae) with Annona macroprophyllata fruits in orchards of Chiapas, Mexico / Interacción de Bephratelloides cubensis (Hymenoptera: Eurytomidae) con frutos de Annona macroprophyllata en huertos de Chiapas, México

Introduction: Annona macroprophyllata Donn. Smith. (Annonaceae) (syn. Annona diversifolia Saff.) is a valued fruit tree species known as papausa. In Mexico and Central America, this fruit has become an important crop because of its tasty flavor and high pulp content. Its fruits are frequently damaged by the incidence of wasps of the genus Bephratelloides Girault (Hymenoptera: Eurytomidae), which develop inside the seeds. Objective: To report the interaction of Bephratelloides cubensis Ashmead during its life cycle in fruits of A. macroprophyllata. Methods: We periodically collected fruits in different states of growth recording a) oviposition, b) the moment of evident infection, c) the development of the wasps inside the seeds, and d) their emergence as adults. We also determined the proportion of damaged fruits and seeds. Results: The data indicate that wasps preferred to oviposit on fruits with a diameter of less than 8 cm, oviposition was more frequent between 11:00 am and 03:00 pm., and there was 26 % infestation of fruits, and 9 % of seeds. Conclusion: It is an obligatory interaction for the wasp, the highest proportion of attack on fruits was in the early stages of fruit development and control actions should focus on the protection of these early stages.

Introducción: Annona macroprophyllata Donn. Smith. (Annonaceae) es una especie de árbol frutal, conocida como papausa. En México y América Central, esta fruta se ha convertido en un cultivo importante y valorado debido a su delicioso sabor y la cremosa textura de su pulpa. Sus frutos son frecuentemente dañados por la incidencia de avispas del género Bephratelloides Girault (Hymenoptera: Eurytomidae), que se desarrollan dentro de las semillas. Objetivo: Describir la interacción de Bephratelloides cubensis Ashmead durante su ciclo de vida en frutos de A. macroprophyllata. Método: Recolectamos frutos en diferentes estados de crecimiento registrando a) la ovoposición, b) el momento de la infección evidente, c) el desarrollo de las avispas dentro de las semillas, y d) su emergencia como adultos. También determinamos la proporción de frutas y semillas dañadas. Resultados: Los datos indican que las avispas preferían ovipositar en frutas con un diámetro de menos de 8 cm, la oviposición era más frecuente entre las 11:00 a.m. y las 3:00 p.m., y que había un 26 % de la infestación en las frutas y un 9 % en semillas. Conclusión: Es una interacción obligada para la avispa y la mayor proporción de ataque a los frutos fue en las etapas tempranas de desarrollo de frutos y las acciones de control se deberían enfocar a la protección de estas etapas.

Chemical composition, larvicidal and cytotoxic activity of Annona salzmannii (Annonaceae) seed oil

The seed oil of Annona salzmannii A. DC. was analyzed by GC-MS and 1H qNMR, revealing a mixture of unsaturated (80.5%) and saturated (18.7%) fatty acids. Linoleic (45.3%) and oleic (33.5%) acid were the major unsaturated fatty acids identified, while palmitic acid (14.3%) was the major saturated fatty acid. The larvicidal effects of A. salzmannii seed oil were evaluated against third-instar larvae of Aedes aegypti (Linn.). The oil exhibited moderate larvicidal activity, with a LC50 of 569.77 ppm (95% CI = 408.11 to 825.88 ppm). However, when the cytotoxic effects of the oil were evaluated, no expressive antiproliferative effects were observed in tumor cell lines B16-F10 (mouse melanoma), HepG2 (human hepatocellular carcinoma), K562 (human chronic myelocytic leukemia), HL-60 (human promyelocytic leukemia), and non-tumor cell line PBMC (peripheral blood mononuclear cells), with IC50 values > 50 µg·mL-1. This is the first study to evaluate the chemical composition, larvicidal and cytotoxic activity of A. salzmannii seed oil

Antiproliferative activity of essential oils from three plants of the Brazilian Cerrado: Campomanesia adamantium (Myrtaceae), Protium ovatum (Burseraceae) and Cardiopetalum calophyllum (Annonaceae) / Atividade antiproliferativa dos óleos essenciais de três plantas do Cerrado brasileiro: Campomanesia adamantium (Myrtaceae), Protium ovatum (Burseraceae) e Cardiopetalum calophyllum (Annonaceae)

Abstract Essential oils, which may be extracted from several parts of plants, have different biological activities. The Brazilian Cerrado has a large variety of plants that yield essential oils, even though many have not been studied yet. Taking into account the biodiversity of this biome, this study aimed at evaluating the antiproliferative activity of essential oils extracted from three species of plants of the Cerrado in Goiás state: Campomanesia adamantium (Cambess.) O. Berg, Protium ovatum (Engl. in Mart.) and Cardiopetalum calophyllum (Schltdl.). Essential oils were extracted from both C. adamantium and C. calophyllum leaves and from P. ovatum leaves and green fruits by hydrodistillation carried out by a Clevenger-type apparatus. The chemical composition of the essential oils was determined by Gas Chromatography coupled to Mass Spectrometry (GC-MS). The following major chemical constituents were identified in the essential oils under investigation: β-myrcene (62.00%), spathulenol (28.78%), germacrene-B (18.27%), β-caryophyllene oxide (16.40%), β-caryophyllene (14.00%), α-pinene (11.30%), viridiflorol (9.99%), limonene (7.30%) and (Z,E)-pharnesol (6.51%). The antiproliferative activity was evaluated in different human tumor cell lines: breast adenocarcinoma (MCF-7), cervical adenocarcinoma (HeLa) and glioblastoma (M059J). A normal human cell line was included (GM07492A, lung fibroblasts). Results showed that essential oils from C. adamantium leaves got the lowest values of IC50 in all strains of tumor cells under evaluation. They were significantly lower than the ones of the normal cell line, an evidence of selectivity. It is worth mentioning that this is the first report of the antiproliferative activity of essential oils from C. adamantium , P. ovatum and C. calophyllum against human tumor cells.

Resumo Os óleos essenciais podem ser extraídos de várias partes das plantas e apresentam diversas atividades biológicas. O Cerrado brasileiro possui uma grande variedade de plantas produtoras de óleos essenciais muitas delas ainda não estudadas. Levando-se em consideração a biodiversidade desse bioma, o objetivo deste trabalho foi avaliar a atividade antiproliferativa dos óleos essenciais extraídos de três espécies de plantas ocorrentes no Cerrado do estado de Goiás: Campomanesia adamantium (Cambess.) O. Berg, Protium ovatum (Engl. in Mart.) e Cardiopetalum calophyllum (Schltdl.). Os óleos essenciais foram obtidos das folhas de C. adamantium e C. calophyllum e das folhas e frutos verdes de P. ovatum por hidrodestilação, usando o aparelho do tipo Clevenger. A composição química dos óleos essenciais foi determinada pelo método de Cromatografia Gasosa acoplada à Espectrometria de Massas (CG-EM). Os constituintes químicos majoritários identificados nos óleos essenciais estudados foram: β-mirceno (62,00%), espatulenol (28,78%), germacreno-B (18,27%), óxido de β-cariofileno (16,40%), β-cariofileno (14,00%), α-pineno (11,30%), viridiflorol (9,99%), limoneno (7,30%) e (Z,E)-farnesol (6,51%). A atividade antiproliferativa foi avaliada em diferentes linhagens de células tumorais humanas: adenocarcinoma de mama (MCF-7), adenocarcinoma cervical (HeLa) e gliobastoma (M059J), além de, uma linhagem celular humana normal (GM07492A, fibroblastos pulmonares). O óleo essencial das folhas de C. adamantium exibiu menores valores de CI50 em todas as linhagens celulares tumorais avaliadas, sendo menores que aquele obtido na linhagem celular normal, indicando seletividade. Este é o primeiro relato da atividade antiproliferativa dos óleos essenciais de C. adamantium , P. ovatum e C. calophyllum contra células tumorais humanas.

Anti-inflammatory, antinociceptive and antioxidant activities of the hydromethanolic fraction from Annona nutans leaves / Atividade anti-inflamatória, antinociceptiva e antioxidante da fração hidrometanólica das folhas de annona nutans

Annona nutans (Annonaceae) is a plant species found in Bolivia, Paraguay, Argentina, and the Brazilian Cerrado, specifically in the states of Mato Grosso and Mato Grosso do Sul (Brazil). Its common names are Araticû-Mi and Araticû-Ñu. The research contributions regarding the chemical composition and biological activities of extracts from A. nutans are rare, with only four articles being found in the literature. Therefore, the present study evaluated the anti-inflammatory and antinociceptive activities of the hydromethanolic fraction (FHMeOH) using carrageenan-induced paw edema and hot-plate tests. In addition, the antioxidant activity was evaluated by DPPH radical scavenging, total phenolic, flavonoid and tannin content assays and quantification of the major metabolites by LC-MS were performed. Oral treatment with the FHMeOH (at a dose of 300 mg.kg-1) significantly reduced paw edema 2 h and 4 h after the inflammatory stimulus. The intraperitoneal (i.p.) treatment with the FHMeOH (50 and 100 mg.kg-1) proved to be most effective, and the inhibition of acute inflammation was still visible 6 h after carrageenan injection. At doses of 50 and 100 mg.kg-1 (i.p.), FHMeOH exhibits central antinociceptive effects by increasing the latency of the reaction in the hot-plate model. The FHMeOH showed antioxidant potential, and the metabolites quercetin-3-O-galactoside, quercetin-3-O-glucoside, isorhamnetin-3-O-galactoside, quercetin-3-O-ß-D-apiofuranosyl-(1→2)-galactopyranoside, and chlorogenic acid were identified and quantified by LC-MS. Our results indicate, for the first time, that FHMeOH has anti-inflammatory, antinociceptive and antioxidant potential, and it is a promising source of studies for new herbal medicines

Annona nutans (Annonaceae) é uma espécie de planta encontrada na Bolívia, Paraguai, Argentina e no Cerrado brasileiro, especificamente nos estados de Mato Grosso e Mato Grosso do Sul (Brasil). Seus nomes mais comuns são aratico e Araticû-Mi e Araticû-Ñu. As contribuições da pesquisa em relação à composição química e atividades biológicas dos extratos de A. nutans são raras, com apenas quatro artigos encontrados na literatura. Portanto, o presente estudo avaliou as atividades anti-inflamatória e antinociceptiva da fração hidrometanólica (FHMeOH) utilizando edema de pata induzido por carragenina e testes de placaquente. Além disso, a atividade antioxidante foi avaliada por meio de sequestro de radical DPPH, e foram realizados ensaios de quantificação de fenóis, flavonoides e taninos totais e quantificação dos principais metabólitos por CL-EM. O tratamento oral com a FHMeOH (na dose de 300 mg.kg-1) reduziu significativamente o edema da pata 2 e 4 h após o estímulo inflamatório. Por outro lado, o tratamento intraperitoneal (i.p.) com FHMeOH (50 e 100 mg.kg-1) provou ser mais eficaz e a inibição da inflamação aguda foi ainda visível 6 horas após a injeção de carragenina. Nas doses de 50 e 100 mg.kg-1 (i.p.), FHMeOH exibiu efeitos antinociceptivos centrais aumentando a latência da reação no modelo de placa quente. FHMeOH apresentou potencial antioxidante e os metabólitos quercetina-3-O-galactosídeo, quercetina-3-O-glicosídeo, isoramnetina-3-O-galactosídeo, quercetina- 3-O-ß-D-apiofuranosil-(1 → 2)-galactopiranosídeo e ácido clorogênico foram identificados e quantificados por CL-EM. Nossos resultados indicam, pela primeira vez, que o FHMeOH possui efeitos anti-inflamatórios, antinociceptivos e antioxidantes, sendo uma fonte promissora de estudos para novos medicamentos fitoterápicos.

Growth and physiology of annona squamosa l. under different irrigation depths and phosphate fertilization / Crescimento e fisiologia de mudas de Annona squamosa L. sob diferentes lâminas de irrigação e adubação fosfatada

This study aimed to evaluate the growth and physiology of Annona squamosa seedlings under increasing irrigation depths and phosphorus doses. The experiment was conducted in protected environment, evaluating five irrigation depths and four P2O5- doses, arranged in 5 x 4 factorial scheme, in randomized blocks, with four replicates and one plant per plot. Treatments were irrigation depths of 60, 80, 100, 120 and 140% of the real evapotranspiration of the seedlings ­ ETr and phosphorus doses of 0, 350, 700 and 1050 mg dm-3 of P2O5-. Plants were evaluated at 120 days after transplanting with respect to growth, gas exchanges, leaf water potential and total dry matter accumulation. Phosphorus dose of 350 mg dm-3 promotes satisfactory dry matter accumulation for A. squamosa seedlings, especially under 100% ETr irrigation. Leaf water potential and gas exchanges of A. squamosa are affected by water stress, through both lack and excess of water. Stomatal conductance is the variable most sensitive to the lack of water, whereas photosynthesis and water use efficiency are the most sensitive to the excess of water. Irrigation depth of 100% ETr is ideal to produce A. squamosa seedlings, but the irrigation depth of 80% ETr can be used to produce these seedlings, disregarding small losses of growth

Objetivou-se avaliar o crescimento e a fisiologia de mudas de A. squamosa sob lâminas crescentes de irrigação e doses de fósforo. O experimento foi conduzido em ambiente protegido, avaliando cinco lâminas de irrigação e quatro doses P2O5, arranjados em esquema fatorial 5 x 4, em blocos casualizados, com quatro repetições e uma planta por parcela. Os tratamentos das lâminas foram 60, 80, 100, 120 e 140% da evapotranspiração real das mudas ­ ETr e as quatro doses de fósforo 0, 350, 700 e 1050 mg dm-3 de P2O5. As plantas foram avaliadas aos 120 dias após o transplantio, quanto ao crescimento, trocas gasosas, potencial hídrico foliar e acúmulo de matéria seca total. A dose de fósforo de 350 mg dm-3 proporciona acúmulo de matéria seca satisfatório para as mudas de A. squamosa, principalmente sob irrigação com 100% da ETr. O potencial hídrico foliar e as trocas gasosas da A. squamosa são afetadas pelo estresse hídrico, tanto por falta como por excesso de água, sendo a condutância estomática a variável mais sensível a falta de água e a fotossíntese e eficiência do uso da água as mais sensíveis ao excesso de água. A lâmina de 100% da ETr é a ideal para a produção de mudas de A. squamosa, porém a lamina de 80% da ETr pode ser utilizada para a produção de mudas de A. squamosa, admitindo-se pequenas perdas no crescimento.

Rollinia mucosa (Jacq) Baillon (Annonaceae) active metabolites as alternative biocontrol agents against the lace bug Corythucha gossypii (Fabricius): an insect pest / Metabolitos activos de la semilla de Rollinia mucosa (Jacq) Baillon (Annonaceae) como agentes alternativos de biocontrol sobre el insecto fitopatógeno Corythucha gossypii (Fabricius) / Metabólitos ativos de Rollinia mucosa (Jacq) Baillon (Annonaceae) como agentes de biocontrole alternativos contra Corythucha gossypii (Fabricius): um inseto-praga

Abstract The lace bug, Corythucha gossypii (Fabricius) is a serious pest affecting over 24 wild and commercially important plant species of the families Annonaceae, Passiflorcae, Caricaceae, Euphorbiaceae, and Solanaceae. Thus far, commercial insecticides, such as 0.1% Dimethoate and 0.1% Imidacloprid have shown effectiveness against this insect, but no botanical pesticides are available to control this bug. In the present study, a Rollinia mucosa (Jacq.) Baillon ethanol extract was evaluated as a biological control agent against the lace bug. Through a toxicity assay involving Artemia salina, the median lethal concentration (LC50) of a raw ethanol extract of R. mucosa seeds was determined, as well as that of its Acetogenin (F1) and Alkaloid (F2) fractions; these LC50 were 0.184, 0.082, and 0.0493 μg/mL, respectively. In addition, with an insecticide assay on lace bug nymphs, a mortality percentage of 86.67% at 5 μg/mL after 72h was observed. These data demonstrate that the R. mucosa seed extract is highly active. Further chemical characterization studies revealed that the main active metabolites contributing to extract activity were acetogenins and alkaloids.

Resumen El hemíptero, Corythucha gossypii (Fabricius) es un insecto que causa daño sustancial en cultivos de más de 24 especies de plantas de las familias Annonaceae, Passifloraceae, Caricaceae, Euphorbiaceae y Solanaceae. En su mayoría estas plantas son de interés económico. Aunque insecticidas comerciales como el Dimetoato (0.1%) y el Imidacloprid (0.1%) permiten un manejo eficiente de este insecto-plaga, no se han reportado alternativas botánicas para estos insecticidas sintéticos. En el presente estudio se evaluó el extracto etanólico de la semilla de Rollinia mucosa (Jacq.) Baillon, como un biocontrolador de C. gossypii. A través de un test de toxicidad con Artemia salina se determinó que la concentración del extracto etanólico letal para el 50% de la población bajo estudio (LC50) fue de 0.184 μg/mL. De igual modo se identificó que las fracciones de acetogeninas (F1) y de alcaloides (F2) de este extracto tienen un LC50 de 0.082 y 0.0493 μg/mL, respectivamente. En el ensayo insecticida con ninfas de C. gossypii se observó una mortalidad del 86.67% después de 72 horas de exposición al extracto etanólico a una concentración de 5 μg/mL. Lo anterior demuestra que el extracto es altamente activo. La caracterización química del extracto evidenció que los principales metabolitos activos que contribuyen a su actividad insecticida son las acetogeninas y los alcaloides.

Resumo Corythucha gossypii (Fabricius) é uma praga séria que afeta mais de 24 plantas silvestre e de interesse comercial, pertencentes as famílias Annonaceae, Passifloraceae, Caricaceae, Euphorbiaceae e Solanaceae. Até o momento, inseticidas comerciais como Dimetoato (0.1%) e Imidacloprid (0.1%) apresentam um controle eficiente sobre este inseto, entretanto não há reportes de pesticidas de origem vegetal para o seu controle. No presente estudo, o extrato etanólico de Rollinia mucosa (Jacq.) Baillon foi avaliado como um controle biológico contra Corythucha gossypii. Por meio do ensaio de toxicidade com Artemia salina a concentração letal média (LC50) para o extrato etanólico das sementes, suas frações de acetogeninas (F1) e fração de alcaloides (F2) foi de 0.184, 0.082 y 0.0493 μg/mL, respetivamente. Adicionalmente, na avaliação do ensaio inseticida, se obteve uma porcentagem de mortalidade de 86.67% à concentração de 5 μg/mL após de 72 horas de exposição, demonstrando uma alta atividade do extrato de sementes de R. mucosa. Os estudos em relação à caracterização química evidenciaram que os principais metabólitos que aportam à atividade do extrato foram as acetogeninas e alcaloides.

Leaf Anatomy of Xylopia aromatica (Lam. ) Mart. (Annonaceae) occurring in a rocky savannah in the Brazilian Amazonian / Anatomia foliar de Xylopia aromatica (Lam. ) Mart. (Annonaceae) ocorrente em uma savana rochosa na Amazônia Brasileira

Savannas of the Amazon Region of Mato Grosso State have an unique ecological identity due to the complexity of the Cerrado-Amazon transition region, their geographical isolation and the physical-chemical properties of the soil. This study aimed to characterize the leaf anatomy of Xylopia aromatica, and to identify potential adaptive traits to the xeric environment. We collected adult leaves from X. aromatica from a Amazonian savannah located in the Nova Canãa do Norte city, Mato Grosso. The leaves were fixed and stored in 70% ethanol. Cross sections were obtained by free hand with the aid of a razor blade, stained with astra blue and basic fuchsin, and mounted on histological slides. Charactersconsidered adaptive: thick cuticle, epidermal cells with thickened walls, the presence of silica in theepidermis, trichomes, hypostomatic leaves, stomata on the same level as the other epidermal cells, presenceof hypodermis, and dorsiventral mesophyll with palisade parenchyma occupying more than 50%. Theanatomical characters presented are of great importance for the establishment and development of X.aromatica in xeric environments. They contribute to the protection of the leaves from many biotic andabiotic factors to which they are subjected, thus ensuring the species survival in the savannic environment.

A região de savana amazônica do Estado do Mato Grosso possui identidade ecológica própria, explicada pela complexa região de transição Cerrado-Amazônia, pelo isolamento geográfico e pelas propriedades físico-químicas do solo. O objetivo deste estudo foi caracterizar a anatomia foliar de Xylopiaaromatica (Lam.) Mart, identificando possíveis características adaptativas ao ambiente xerófilo. Foram coletadas folhas adultas de X. aromatica em savana amazônica, localizado no município de Nova Canaã do Norte - Mato Grosso. As folhas foram fixadas em FAA50 e armazenadas em etanol a 70%. Os cortes transversais foram obtidos a mão livre com auxílio de lâmina de barbear, corados com azul de Astra e fucsina básica e montados em lâminas histológicas. Caracteres aqui considerados adaptativos: cutículaespessa, células epidérmicas com paredes espessadas, presença de sílica na epiderme, tricomas, folhashipoestomáticas, estômatos no mesmo nível das demais células epidérmicas, presença de hipoderme, mesofilo dorsiventral com parênquima paliçádico ocupando mais de 50% e lacunoso bem compacto. Os caracteres anatômicos aqui apresentados são de grande importância para as espécies desenvolverem-se em ambientes xerófilos, demonstrando contribuição para a proteção das folhas a diversos fatores bióticos e abióticos aos quais esta se sujeita, garantindo a sobrevivência das espécies no ambiente savânico.

Preliminary analysis of Brazilian Amazonian plant extracts using bench-top assays in a through-put basis aiming the identification of potential antioxidant natural product

Plants are a source of compounds that are used for the treatment of human and veterinary diseases. Brazil is one of the richest countries in the world in terms of biodiversity. The present study evaluated extracts using thin-layer chromatography to identify antioxidant activity and determine the presence of groups of compounds, in a large-scale basis. A total of 1,260 aqueous and organic extracts were obtained from plants that were collected in the Amazon and Atlantic rain forests. Thin-layer chromatography was performed to evaluate the presence of alkaloids, anthraquinones, cardioactive glicosides, terpenes, and phenolics and determine antioxidant and radical scavenging activity using the following reagents: Dragendorff's reagent, KOH, Kedde's reagent, sulphuric acid, NP reagent, ß-carotene, and 2,2-diphenyl-1-picrylhydrazyl (DPPH). Of the 1,260 extracts, 837 (66.43%) presented a ß-carotene/bleaching response, and 1,205 (95.63%) presented a radical scavenging response. Alkaloids were found in 203 extracts (16.11%). Anthraquinones were found in 14 extracts (1.11%), cardenolides were found in eight extracts (0.63%). The present findings shows the importance of the Brazilian Amazon plants as sources of antioxidant and radical scavenging active compounds.(AU)

Efecto del extracto etanólico de las hojas de Annona muricata L (guanábana) sobre Síndrome metabólico inducido en ratas / Effect of ethanolic extract of the leaves of Annona muricata L (soursop) on metabolic syndrome induced in rats

Objetivo: Evaluar el efecto del extracto etanólico de hojas de guanábana (EEA) administrado en ratas con síndrome metabólico experimental. Materiales y Métodos: Estudio Experimental. 90 ratas machos de 2 meses, cepa Holtzmann de 175±25g de peso fueron divididos en nueve grupos: I) normal (Suero salino fisiológico-SSF-2mL/kg); II) EEA 200mg/kg; III) 200mg/kg de colesterol y 100mg/kg de fructosa (CF) para inducción de síndrome metabólico (SM); IV-VI) CF + EEA 50, 100 y 200mg/kg respectivamente; VII) CF + atorvastatina 20mg/kg; VIII) CF + enalapril 20mg/kg; IX) CF + enalapril 20mg/kg + atorvastatina 20mg/kg. corporal. Estos grupos fueron intervenidos por 90 días, donde se midieron: peso corporal (g), niveles de glicemia (mg/dL), hemoglobina glicosilada (HbA1c), presión arterial (mmHg) y perfil lipídico (Colesterol total, triglicéridos, HDL-c y LDL-c en mg/dL). Resultados: Se encontraron diferencias significativas en comparación, en los valores de glicemia (-23.9 mg/ml; IC95%: -36.7 a -11.0), HbA1c (-0.8%; IC95%: -1.2 a -0.4) y Triglicéridos (-68.1; IC95% -96.0 a -40.1) en la dosis de EEAM 200mg/kg en comparación con el grupo III. La dosis de 100mg/kg logró presiones arteriales: sistólica (-6.8 mmHg; IC95%: -17.5 a 3.9, p=0.397), diastólica (-3.9 mmHg; IC95%: -14.0 a 6.2, p=0.861) y media (-9.9 mmHg; IC95%: -31.1 a 11.3, p=0.720); parecidas al grupo I. Asimismo, esta dosis logró diferencias en colesterol total (-57.6 mg/dl; IC 95% -75.7 a -39.4) y LDL-c (-56.5 mg/dl; IC 95% -75.7 a -37.3) en comparación con el grupo III. Conclusiones: El extracto etanólico de las hojas de Annona muricata L. (guanábana) a dosis de 200 mg/Kg mostró un mejor efecto ante la inducción de síndrome metabólico experimental.

Avaliação da atividade anti-Leishmania e anti-Trypanosoma do extrato etanólico das folhas de Annona squamosa L / Evaluation of the anti-leishmania and anti-trypanosoma activity of ethanolic extract from the leaves of Annona squamosa L

As doenças parasitárias, também chamadas de “doenças negligenciadas”, continuam sendo uma grande dificuldade para o desenvolvimento social e econômico dos países mais pobres. Podemos citar como exemplo dessas doenças, a leishmaniose e a doença de Chagas. A leishmaniose é causada por parasitas do gênero Leishmania e afeta cerca de 12 milhões de pessoas. A doença de Chagas, causada pelo protozoário Trypanosoma cruzi, causa aproximadamente 50.000 mortes por ano. Os fármacos disponíveis para o tratamento dessas doenças são altamente tóxicos, sendo este um dos motivos que leva à busca por drogas eficazes e seguras para seus tratamentos. As folhas da Annona squamosa, espécie da família Annonaceae, já foram descritas na literatura por suas atividades hepatoprotetora, antiparasitária, pesticida e antimicrobiana. Nesse estudo avaliamos a atividade anti-leishmania e tripanocida do extrato etanólico das folhas de Annona squamosa L. (EEAS) em formas promastigota do parasita Leishmania braziliensis e Leishmania infantum e epimastigota de Trypanosoma cruzi, além de avaliar a atividade citotóxica em fibroblasto. Os resultados demonstram que o extrato apresentou uma melhor atividade contra Leishmania infantum e Leishmania brasiliensis quando comparados com Trypanosoma cruzi; e que apresentou uma maior toxicidade nas concentrações de 500 e 1000 μg/ml, com mortalidade dos fibroblastos de aproximadamente 85% e 100%, respectivamente. Esse estudo aponta para uma perspectiva terapêutica alternativa que se mostrou eficaz frente aos parasitas aqui estudados, exceto a forma epimastigota de Trypanosoma cruzi. Com relação aos testes de citotoxicidade fazem-se necessários novos testes, uma vez que apresentou um alto nível de toxicidade, viabilizando assim futuros ensaios in vivo.

The parasitic diseases, also calls by “neglected diseases”, continue being a major difficulty for the social and economic development of the poorest countries. We can cite as an example of these diseases, the leishmaniasis and the Chagas disease. Leishmaniasis is caused by parasites of the genus Leishmania and affects about 12 million people. The Chagas disease, caused by the protozoan Trypanosoma cruzi, causes approximately 50,000 deaths per year. The drugs available for the treatment of these diseases are highly toxic, being this one of the reasons that leads to the search for effective and safe drugs for their treatments. The leaves of the Annona squamosa, species of the family Annonaceae, have already been described in the literature by their hepatoprotective activities, antiparasitic, pesticide and antimicrobial. In this study we assessed the activity tripanocidal and antileishmania of ethanolic extract from the leaves of Annona squamosa L. (EEAS) in promastigota forms of the parasite Leishmania braziliensis and Leishmania infantum and epimastigota of Trypanosoma cruzi, in addition to evaluating the cytotoxic activity in fibroblasts. The results demonstrate that the extract presented a better activity against Leishmania infantum and Leishmania brasiliensis when compared with Trypanosoma cruzi; and which presented a greater toxicity at concentrations of 500 and 1000 μg/ml, with mortality of fibroblasts of approximately 85% and 100%, respectively. This study points to an alternative therapeutic perspective that showed effective against the parasites here studied, except the epimastigota form of Trypanosoma cruzi. With relation to cytotoxicity tests are required new tests, once presented a high level of toxicity, thus enabling future in vivo assays.

Searching for promising sources of grain protectors in extracts from neotropical annonaceae / Búsqueda de fuentes prometedoras de protectores de granos en extractos de anonaceas neotropicales

To investigate potential sources of novel grain protector compounds against Sitophilus zeamais (Coleoptera: Curculionidae), which is an important insect pest of stored cereals, this study evaluated the bioactivity of ethanolic extracts (66) prepared from 29 species belonging to 11 different genera of Neotropical Annonaceae. A screening assay demonstrated that the most pronounced bioactive effects on S. zeamais were caused by ethanolic extracts from Annona montana, A. mucosa, A. muricata, and A. sylvatica seeds, causing the death of all weevils exposed, almost complete inhibition of the F1 progeny and a drastic reduction in grain losses. Furthermore, the ethanolic extracts obtained from the leaves of A. montana, A. mucosa, A. muricata, and Duguetia lanceolata, especially A. montana and A. mucosa, demonstrated significant bioactive effects on the studied variables; however, the activity levels were less pronounced than in the seed extracts, and the response was dependent on the concentration used. This study is the first to report the activity of secondary metabolites from D. lanceolata on insects as well as the action of A. sylvatica on pests associated with stored grains.

Para investigar las posibles fuentes de nuevos compuestos protectores de granos contra Sitophilus zeamais (Coleoptera: Curculionidae), una importante plaga de los cereales almacenados, este estudio evaluó la bioactividad de los extractos etanólicos (66) preparados a partir de 29 especies pertenecientes a 11 géneros distintos de Anonaceas Neotropicales. Un ensayo de selección demostró que los efectos bioactivos más relevantes sobre S. zeamais fueron causados por los extractos etanólicos de las semillas de Annona montana, de A. mucosa, de A. muricata y de A. sylvatica, que causaron la muerte de todos los gorgojos expuestos, la inhibición parcial de la progenie F1 y una drástica reducción de las pérdidas de grano. Además, los extractos etanólicos obtenidos de las hojas de A. montana, de A. mucosa, de A. muricata y de Duguetia lanceolata, especialmente de A. montana y de A. mucosa, demostraron efectos bioactivos significativos sobre las variables estudiadas. Sin embargo, los niveles de bioactividad fueron menores en comparación con los extractos de semillas, y la respuesta fue dependiente de la concentración utilizada. Este estudio es el primer relato sobre la actividad de los metabolitos secundarios de D. lanceolata sobre insectos, así como la acción de A. sylvatica sobre plagas asociadas a los granos almacenados.

Potential phytoinsecticide of Annona mucosa (Jacq) (Annonaceae) in the control of brown stink bug / Potencial fitoinseticida de Annona mucosa (Jacq) (Annonaceae) no controle do percevejo-marrom

Plant extract have been recognized as important natural source of insecticide. Herein, we assessed the toxicity of leaves-extract and seeds-extract of Annona mucosa (Annonaceae) against brown stink bug, Euschistus heros (F.) (Hemiptera: Pentatomidae), a soybean pest in Neotropical America. For this, bioassays with nymphs and adults were conducted using the concentrations of 5, 10, 20, 40 and 80 mg.mL-1 of leaves and seeds extract of A. mucosa and DMSO (10%), as control. Ten replicates were conducted by treatment, with five nymphs (3rd instar) by replicate (bioassay with nymphs) and one couple of adults (< 24h of old) by replicate (bioassay with adults). The number of nymphs death (mortality) and survival of adults were assessed. For adults the fecundity (number of eggs) and fertility (number of nymphs hatched) of females also were assessed. The nymphs bioassay has promising results in seeds extract, when compared to leaves extract. The mortality of nymphs treated with seeds extract was dependent-concentration, with increased in mortality in concentrations > 5.0mg.mL-1, while nymphs treated with leaves extract do not exhibit difference between treatments. In adults the survival was reduced, when were treated with seed extract of A. mucosa at concentrations higher than 20mg.mL-1, with reduction up to 92 days in contrast to control. A drastic reduction in the fecundity and fertility was observed, which highlight deleterious action of seed extract against reproductive potential of E. heros. This is the first report about toxicity A. mucosa (leaves and seed) on E. heros, and our results suggest that seeds extract of A. mucosa is promising for control of nymphs and adults of brown stink bug.

Extratos de plantas tem sido reconhecidos como um importante recurso natural de inseticidas. Nesta pesquisa avaliou-se a toxicidade do extrato de folhas e de sementes de Annona mucosa (Annonaceae) sobre o percevejo marrom, Euschistus heros (F.) (Hemiptera: Pentatomidae), uma praga-chave na cultura da soja na America Neotropical. Para tanto, bioensaios com ninfas e adultos foram conduzidos usando as concentrações de 5, 10, 20, 40 e 80 mg.mL-1 do extrato de folhas e sementes de A. mucosa e DMSO (10%), como controle. Dez repetições foram conduzidas por tratamento, com cinco ninfas (3º instar) por repetição (bioensaio com ninfas) e um casal de adultos (< 24h de idade) por repetição (bioensaio com adultos). Foram avaliados o número de ninfas mortas (mortalidade) e a sobrevivência dos adultos. Para fêmeas também foi avaliada a fecundidade (número de ovos) e fertilidade (número de ninfas eclodidas). O extrato de semente de A. mucosa apresentou resultados promissores na mortalidade de ninfas de E. heros, quando comparado ao extrato de folhas. A mortalidade de ninfas tratadas com extrato de semente foi dependente da concentração, com aumento da mortalidade em concentrações > 5,0mg.mL-1, enquanto as ninfas tratadas com extrato de folhas não apresentaram diferença entre os tratamentos. A sobrevivência dos adultos foi reduzida, quando estes foram tratados com extrato de semente de A. mucosa em concentrações maiores que 20mg.mL-1, com redução de até 92 dias, em contraste ao controle. Foi observada uma drástica redução na fecundidade e fertilidade, o que destaca a ação deletéria do extrato de sementes sobre o potencial reprodutivo de E. heros. Este é o primeiro registro sobre a toxicidade de A. mucosa (folhas e sementes) sobre E. heros e os resultados sugerem que o extrato de semente de A. mucosa é promissor para o controle de ninfas e adultos do percevejo marrom.

Pseudoxandra sclerocarpa maas, colombian medicinal plant: a review / Pseudoxandra sclerocarpa maas, planta medicinal colombiana: una revisión

The Annonaceae family is one of the largest, with 130 genre and 2500 species, consisting of trees, shrubs and a few vines. Within this family, the Pseudoxandra genus of neotropical distribution is found. In Colombia, there are endemic and native species, distributed in the Amazon region and along the valley of the Magdalena River in the Chocó. Of the total of 22 species that form the genus in Colombia, Pseudoxandra sclerocarpa Maas is exclusive of the Antioquia region, receiving the common name of garrapato or frisolo. From this tree, bisbenzylisoquinolinic alkaloids like antioquine, medellline, obaberine, among others, has been isolated. Also neolignans: dieugenol and dehydro-1-O-methydehydrodieugenol. It is used in traditional medicine as an antiparasitic. From the pharmacological point of view, the alkaloids have shown activity against leishmaniasis and also have spasmolytic activity. Within the bisbenzylisoquinolinic alkaloids, antioquine has properties of being calcium antagonist.

La familia Annonaceae es uno de los más grandes, con 130 géneros y 2.500 especies, que consiste en árboles, arbustos y algunos bejucos. Dentro de esta familia, el género Pseudoxandra es de distribución neotropical. En Colombia existen especies endémicas y nativas, distribuidas en la región del Amazonas ya lo largo del valle del río Magdalena en el Chocó. Del total de 22 especies que forman el género en Colombia, Pseudoxandra sclerocarpa es exclusiva de la región de Antioquia, recibiendo el nombre común de garrapato o frisolo. De este árbol se ha aislado alcaloides bisbencilisoquinolínicos como antioquina, medelllina, obaberina, entre otros. También neolignanos: dieugenol y dehidro-1-O-metil-dehidrodieugenol. Se utiliza en la medicina tradicional como un antiparasitario. Desde el punto de vista farmacológico, los alcaloides han mostrado actividad contra la leishmaniasis y también tienen actividad espasmolítica. Dentro de los alcaloides bisbencilisoquinolínicos, antioquina tiene propiedades de ser antagonista del calcio.

In vitro biological screening of the anticholinesterase and antiproliferative activities of medicinal plants belonging to Annonaceae

The aim of this research was to investigate the antiproliferative and anticholinesterase activities of 11 extracts from 5 Annonaceae species in vitro. Antiproliferative activity was assessed using 10 human cancer cell lines. Thin-layer chromatography and a microplate assay were used to screen the extracts for acetylcholinesterase (AchE) inhibitors using Ellman's reagent. The chemical compositions of the active extracts were investigated using high performance liquid chromatography. Eleven extracts obtained from five Annonaceae plant species were active and were particularly effective against the UA251, NCI-470 lung, HT-29, NCI/ADR, and K-562 cell lines with growth inhibition (GI50) values of 0.04-0.06, 0.02-0.50, 0.01-0.12, 0.10-0.27, and 0.02-0.04 µg/mL, respectively. In addition, the Annona crassiflora and A. coriacea seed extracts were the most active among the tested extracts and the most effective against the tumor cell lines, with GI50 values below 8.90 µg/mL. The A. cacans extract displayed the lowest activity. Based on the microplate assay, the percent AchE inhibition of the extracts ranged from 12 to 52%, and the A. coriacea seed extract resulted in the greatest inhibition (52%). Caffeic acid, sinapic acid, and rutin were present at higher concentrations in the A. crassiflora seed samples. The A. coriacea seeds contained ferulic and sinapic acid. Overall, the results indicated that A. crassiflora and A. coriacea extracts have antiproliferative and anticholinesterase properties, which opens up new possibilities for alternative pharmacotherapy drugs.

In vitro antioxidant properties, free radicals scavenging activities of extracts and polyphenol composition of a non-timber forest product used as spice: Monodora myristica

BACKGROUND: Excessive production of free radicals causes direct damage to biological molecules such as DNA, proteins, lipids, carbohydrates leading to tumor development and progression. Natural antioxidant molecules from phytochemicals of plant origin may directly inhibit either their production or limit their propagation or destroy them to protect the system. In the present study, Monodora myristica a non-timber forest product consumed in Cameroon as spice was screened for its free radical scavenging properties, antioxidant and enzymes protective activities. Its phenolic compound profile was also realized by HPLC. RESULTS: This study demonstrated that M. myristica has scavenging properties against DPPH',OH',NO', and ABTS'radicals which vary in a dose depending manner. It also showed an antioxidant potential that was comparable with that of Butylated Hydroxytoluene (BHT) and vitamin C used as standard. The aqueous ethanol extract of M. myristica barks (AEH); showed a significantly higher content in polyphenolic compounds (21.44 ± 0.24 mg caffeic acid/g dried extract) and flavonoid (5.69 ± 0.07 quercetin equivalent mg/g of dried weight) as compared to the other studied extracts. The HPLC analysis of the barks and leaves revealed the presence of several polyphenols. The acids (3,4-OH-benzoic, caffeic, gallic, O- and P- coumaric, syringic, vanillic), alcohols (tyrosol and OH-tyrosol), theobromine, quercetin, rutin, catechine and apigenin were the identified and quantified polyphenols. All the tested extracts demonstrated a high protective potential on the superoxide dismutase (SOD), catalase and peroxidase activities. CONCLUSION: Finally, the different extracts from M. myristica and specifically the aqueous ethanol extract reveal several properties such as higher free radical scavenging properties, significant antioxidant capacities and protective potential effects on liver enzymes.

Flavonoid-rich fraction of the Monodora tenuifolia seed extract attenuates behavioural alterations and oxidative damage in forced-swim stressed rats / 中国天然药物

The antidepressant effects of the flavonoid-rich fraction of Monodora tenuifolia seed extract were examined by assessing the extent of attenuation of behavioural alterations and oxidative damage in the rats that were stressed by forced swim test. Compared with the model control group, the altered behavioural parameters were attenuated significantly (P < 0.05) in the group treated with the flavonoid-rich fraction (100 and 200 mg·kg(-1)), comparable to the group treated with the standard drug, fluoxetine (10 mg·kg(-1)). The flavonoid-rich fraction and fluoxetine improved significantly (P < 0.05) the activities of the antioxidant enzymes such as superoxide dismutase and catalase as well as other biochemical parameters such as reduced glutathione, protein, and nitrite in the brain of the stressed rats. These results suggested that the flavonoid-rich fraction of Monodora tenuifolia seed extract exerted the antidepressant-like effects which could be useful in the management of stress induced disease.

Caracterização física e físico-química de frutos de araticum (Annona crassiflora Mart. ) / Physical and physical and chemical characterization of araticum fruits (Annona crassiflora Mart. )

Este trabalho objetivou caracterizar física e quimicamente frutos de araticum coletados de populações naturais de plantas localizadas em cinco áreas de Cerrado do estado de Goiás. O trabalho foi realizado na Escola de Agronomia e Engenharia de Alimentos da Universidade Federal de Goiás. Os frutos maduros foram avaliados individualmente, por área e por planta. Determinaram-se: massas do fruto, da polpa e da casca, diâmetro e altura do fruto, número total de sementes e de sementes defeituosas por fruto, massa das sementes, cores do fruto, da casca e da polpa, textura da casca e teor de sólidos solúveis totais. As características físicas mostraram variação significativa entre frutos de plantas das diferentes áreas. A massa média do fruto foi de 872,06 g. A polpa, parte de importância econômica, representou cerca de 47% dessa massa, cuja variação por região foi de 44% a 49%. O número de sementes foi elevado, em média 108, representando 15,8% da massa do fruto. Há tendência para a cor marrom da casca, assim como para a cor amarelada ou creme da polpa. Observou-se uma variação de textura da casca, desde lisa até rugosa. Do total de 219 frutos avaliados, 15,5% apresentaram o orifício de saída de Bephratelloides pomorum. Houve diferenças significativas entre áreas e entre plantas dentro de áreas para todas as variáveis físicas. A massa média dos frutos de araticum foi de 872,06 g com rendimento médio de polpa, casca e sementes de: 46,8%, 37,4% e 15,8%, respectivamente.

This work objectified to characterize physics chemically and collected fruits of araticum of natural populations of plants located in five areas of Cerrado of the state of Goiás. The work was carried through in the "Escola de Agronomia e Engenharia de Alimentos, Universidade Federal de Goiás". The mature fruits had been evaluated individually, for area and plant. They had been determined: masses of the fruit, the pulp and the rind, diameter and height of the fruit, total number of seeds and defective seeds for fruit, mass of the seeds, colors of the fruit, the rind and the pulp, texture of the rind and total soluble solids. The physical characteristics had shown significant variation between fruits of plants of the different areas. The average mass of the fruit was of 872,06 g. The pulp, part of economic importance, represented about 47% of this mass, whose variation for region was of 44% 49%. The number of seeds was raised, on average 108, representing 15.8% of the mass of the fruit. It has trend for the brown color of the rind, as well as for the yellowish color or cream of the pulp. A variation of texture of the rind was observed, since smooth until rough. Of the total 219 of evaluated fruits, 15.5% had presented the orifice of exit of Bephratelloides pomorum. It inside had significant differences between areas and plants of areas for all the variable physicist. The mass average of the 872,06 fruits of araticum was of g with average income of pulp, rind and seeds of: 46.8%, 37.4% and 15.8%, respectively.

Biología y hábitos del barrenador de la semilla Bephratelloides cubensis ashmead (Hymenoptera: Eurytomidae) / Biology and behavior of the seed borer wasp Bephratelloides cubensis ashmead (Hymenoptera: Eurytomidae)

The soursop Annona muricata is an important fruit for national market, and for exportation, but the crop is affected by pests and diseases. The seed borer wasp Bephratelloides cubensis Ashmead is the pest that produces the highest damage to the crop in Mexico. Sixty percent of damaged fruits and 5-50 seeds per fruit have been registered, with 25 percent reduction in yield. In Nayarit, Mexico, 100 percent of damaged fruits were recorded. In this State, an experiment with soursop was conducted to study the life cycle under field conditions and to determine diurnal behavior of the female of B. cubensis. The highest activity of the wasp was observed between 12:00h and 13:00h (35ºC, 54 percent RH and 409.34 luxes). Females oviposited in fruits with a diameter of 3.1-7.6 cm. Larvae of B. cubensis developed five instars, adults survived no longer than 22 days, and female survived longer than males; they lived 22 and 15 days, respectively. Life cycle of B. cubensis varied from 69 to 122 days.