Abstract Background: Cryptoccocal meningitis continues to present high incidence among AIDS patients. The treatment of choice is the synergistic combination of flucytosine (5-FC) with amphotericin B deoxycholate (AmBd) or its lipid formulations. However, 5-FC is unavailable in many countries and AmB demands hospitalization. The combination of AmB with the fungistatic fluconazole (FLC) or the use of high FLC daily doses alone became the choice. Nonetheless, sterilization of cerebrospinal fluid is delayed with FLC monotherapy, mainly with high fungal burden. These findings suggest the search for new antifungal compounds, such as liriodenine. Methods: Liriodenine antifungal activity was evaluated by three procedures: determining the minimum inhibitory concentration (MIC) on 30 strains of the Cryptococcus neoformans (C. neoformans) complex and 30 of the Cryptococcus gattii (C. gattii) complex, using EUCAST methodology and amphotericin B deoxycholate as control; performing the time-kill methodology in two strains of the C. neoformans complex and one of the C. gattii complex; and injury to cryptococcal cells, evaluated by transmission electron microscopy (TEM). Liriodenine absorption and safety at 0.75 and 1.50 mg.kg-1 doses were evaluated in BALB/c mice. Results: Liriodenine MICs ranged from 3.9 to 62.5 μg.mL-1 for both species complexes, with no differences between them. Time-kill methodology confirmed its concentration-dependent fungicidal effect, killing all the strains below the limit of detection (33 CFU.mL-1) at the highest liriodenine concentration (32-fold MIC), with predominant activity during the first 48 hours. Liriodenine induced severe Cryptococcus alterations - cytoplasm with intense rarefaction and/or degradation, injury of organelles, and presence of vacuoles. Liriodenine was better absorbed at lower doses, with no histopathological alterations on the digestive tract. Conclusion: The fungicidal activity confirmed by time-kill methodology, the intense Cryptococcus injury observed by TEM, the absorption after gavage administration, and the safety at the tested doses indicate that the liriodenine molecule is a promising drug lead for development of anticryptococcal agents.

Resumen 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.

Abstract 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.

Abstract The genus Bursera is a characteristic component of the tropical dry forest (TDF) of Mexico, however, despite being relevant for the conservation and management of the species, its biology is little known, among them, their phenological patterns and their associations with seasonal factors. The aim of this research was to characterize the phenology of Bursera simaruba Jacq. ex L and Bursera tomentosa (Jacq.) Triana & Planch and to evaluate their relationship with environmental variables. The phenological study was realized in a protected area with tropical dry forest of the Central Depresion from Chiapas. Fifty adult trees of each specie were selected and the intensity of four reproductive phenophases (flower bud, open flower, fruit development and permanence of fruits) and three vegetative ones (leaf bud, mature leaves, senescence) were recorded using the Fournier method. Seasonality and synchrony in each phase were estimated by circular statistics calculating the mean vector (r) and the Rayleight test (Z). Also, the functional types of vegetative phenology were categorized and that of reproductive phenology. All the phenological stages were seasonal (r> 0.5), in both species, the flowering and leaf bud phenophases occurred in a short period while the vegetative and fruit production phenophases are the more extended stages. The reproductive and vegetative phenology of Bursera simaruba and B. tomentosa were very similar, and the most correlated environmental variables with them (rs ≥ 0.6) were temperature and humidity. These results allow to know the phenological dynamic of Bursera species in the TDF, and with this, to promote management and conservation strategies of these important species.

Resumen El género Bursera es un componente característico de los bosques tropicales secos (BTS) de México y relevante para la conservación de estos hábitats, sin embargo, sus aspectos fenológicos son poco conocidos. Este estudio tuvo como objetivo caracterizar la fenología de Bursera simaruba Jacq. ex L y Bursera tomentosa (Jacq.) Triana & Planch y evaluar su relación con las variables ambientales. El seguimiento fenológico se realizó en una zona protegida del BTS de la región florística conocida como Depresión Central en Chiapas, México durante un ciclo anual (mayo 2017- abril 2018). Se seleccionaron 50 individuos adultos de cada especie y se registró la intensidad de sus fenofases reproductivas (botón floral, flor abierta, desarrollo de frutos y permanencia de frutos) y fenofases vegetativas (brote foliar, hojas maduras, senescencia) utilizando los métodos de Fournier. La estacionalidad y sincronía de cada fase se estimó mediante estadística circular calculando el vector medio (r) y la prueba Rayleigh (Z). Además, se categorizaron los tipos funcionales de su fenología vegetativa y reproductiva. Las etapas fenológicas en ambas especies fueron estacionales (r > 0.5), la floración y brote foliar sucedieron en un periodo breve mientras que el desarrollo vegetativo y las de producción de frutos fueron etapas más extendidas. Las fenofases reproductiva y vegetativa de Bursera simaruba y B. tomentosa son similares, las variables ambientales más correlacionadas (rs ≥ 0.6) a ellas fueron la temperatura y la humedad. El estudio de estas burseras permite conocer su dinámica en el BTS y con esto proponer planes de manejo de estas especies resiníferas.

Abstract Background: Endemic systemic mycoses remain a health challenge, since these opportunistic diseases are increasingly infecting immunosuppressed patients. The simultaneous use of antifungal compounds and other drugs to treat infectious or non-infectious diseases has led to several interactions and undesirable effects. Thus, new antifungal compounds should be investigated. The present study aimed to evaluate the activity of liriodenine extracted from Annona macroprophyllata on agents of systemic mycoses, with emphasis on the genus Paracoccidioides. Methods: The minimum inhibitory concentration (MIC) and minimum fungicide concentration (MFC) were determined by the microdilution method. The cellular alterations caused by liriodenine on a standard P. brasiliensis (Pb18) strain were evaluated by transmission and scanning electron microscopy. Results: Liriodenine was effective only in 3 of the 8 strains of the genus Paracoccidioides and in the Histoplasma capsulatum strain, in a very low concentration (MIC of 1.95 μg.mL-1); on yeasts of Candida spp. (MIC of 125 to 250 μg.mL-1), including C. krusei (250 μg.mL-1), which has intrinsic resistance to fluconazole; and in Cryptococcus neoformans and Cryptococcus gattii (MIC of 62.5 μg.mL-1). However, liriodenine was not effective against Aspergillus fumigatus at the studied concentrations. Liriodenine exhibited fungicidal activity against all standard strains and clinical isolates that showed to be susceptible by in vitro tests. Electron microscopy revealed cytoplasmic alterations and damage to the cell wall of P. brasiliensis (Pb18). Conclusion: Our results indicate that liriodenine is a promising fungicidal compound that should undergo further investigation with some chemical modifications.

Resumo A presente revisão tem por objetivo apresentar os avanços nos estudos sobre a propagação de espécies da família Annonaceae, que inclui espécies de importância econômica como: graviola, fruta-do-conde, atemoia e cherimoia. Na propagação sexuada, os avanços estão relacionados, principalmente, com a melhor compreensão dos estádios de desenvolvimento das sementes, mecanismos de dormência, e germinação , em especial a fase de embebição das mesmas. Na propagação assexuada, são apresentados estudos de compatibilidade entre enxertos e porta-enxertos, com foco na expressão de genes envolvidos com a formação dos tecidos. Também é discutido o método da estaquia, outra opção de propagação para este grupo de plantas consideradas de difícil enraizamento, abordando-se fatores endógenos e exógenos relacionados ao tema, bem como as estratégias de manejo que interferem no sucesso desta técnica.

Abstract This review aims to present advances in studies on the propagation of the Annonaceae species, which includes species of economic importance such as: soursop, custard apple, atemoya and cherimoya. In sexual propagation, advances are mainly related to a better understanding of the stages of seed development, dormancy mechanisms, and germination. In asexual propagation, compatibility studies between grafts and rootstocks are presented, focusing on the expression of genes involved in tissue formation. The cutting method is also discussed, which is another option for the propagation for this group of plants considered difficult to root, approaching endogenous and exogenous factors related to the subject, as well as management strategies that affect the success of this technique.

Abstract: Background. Phenological descriptions of plant species are used to understand cycles of foliation, flowering and fruiting. These studies provide useful means for understanding the relationships between plants and their environment. Question. What are the the foliar and reproductive strategies of four species of Annona living in the Deciduous Forest of Chiapas, Mexico? Species studied. Annona lutescens, A. macroprophyllata, A. reticulata and A. purpurea. Study sites. Three areas of deciduous forest were studied (16°31’36’’ N, 92°58’25’’ W, 16°28’82’’ N, 92°57’48’’ W, and 16°21’47’’ N, 92°58’32’’ W), located between the towns of Parral and Chiapa de Corzo, Chiapas, Mexico. Method. Stages of foliation, flowering, fruiting and senescence of 20 individuals per species were documented by written and photographic records during the course of a year. Variable time periods were adapted to the extended BBCH scale. The phenology was correlated with the stages of rain and drought by Spearman’s correlations Results. Phenological characteristics distinguish A. reticulata and A. lutescens as species with a reproductive strategy (flowering and fruiting) synchronized with the dry months, in contrast to A. purpurea and A. macroprophyllata, which show a reproductive strategy associated with the rainy season. The foliation of the four species is phenologically related to the rainy season. Conclusions. The species studied could be grouped according to their leaf pattern as brevideciduous or deciduous and according to their reproductive pattern as having long-term or short-term strategies.

Resumen Antecedentes. Las descripciones fenológicas de las especies vegetales son usadas para conocer los ciclos de foliación, floración y fructificación de las plantas cultivadas, estos estudios proveen herramientas útiles para comprender las relaciones entre las plantas y su ambiente. Preguntas. ¿Cuáles serían las estrategias foliares y reproductivas de cuatro especies del género Annona que habitan en la Selva Baja Caducifolia de Chiapas, México? Especies en estudio. Annona lutescens, A. macroprophyllata, A. reticulata y A. purpurea Sitio de estudio. Se estudiaron tres áreas de la selva baja caducifolia, localizada entre los municipios de Chiapas de Corzo y Parral, en Chiapas, México (16° 31’ 36’’ N, 92° 58’ 25’’ W; 16° 28’ 82’ ‘N, 92° 57’ 48’’ W, and 16° 21’ 47’’ N, 92° 58’ 32’’ W). Métodos. Las etapas de foliación, floración, fructificación y senescencia de 20 individuos por especie se documentaron mediante registros fotográficos y escritos durante un año y en periodos de tiempo variable y se adecuaron a la escala BBCH extendida. La fenología y las etapas de lluvia y sequía se asociaron a través de correlaciones de Spearman. Resultados. Las características fenológicas distinguen a A. reticulata y A. lutescens como especies con estrategia de reproducción (floración y fructificación) sincronizadas con los meses de secas, contrastando con las de A. purpurea y A. macroprophyllata que están asociadas a la época de lluvias. La fenología de foliación de las cuatro especies está relacionada con la época de lluvias. Conclusiones. Las especies estudiadas pudieran agruparse de acuerdo a su patrón foliar en brevideciduas y deciduas y de acuerdo a su patrón reproductivo en especies con estrategias largas y cortas

Annona macroprophyllata (papausa or ilama) is an economically important species of fruit that is consumed in large quantities in Central America and is considered to have the finest flavour of all of the custard apples (Annonaceae). This species presents propagation problems including the six-month dormancy of the seeds, which has not been fully explained. Among the factors affecting the germination of Annona seeds are the position inside of the fruit and the presence of a micropylar woody plug. The present work addresses the importance of these factors in the germination and dormancy of this species, using a randomised design with seeds from 169 fruits of the white variety of Annona macroprophyllata. The fruits were divided into three sections (basal, middle, and apical) in which seeds were evaluated for nine months with the following parameters: the size and weight of the seeds and the viability and germination percentage considering the presence or absence of the micropylar plug. The results show no correlation between the breaking of dormancy and either the position of the seeds in the fruit or the micropylar plug, but they do establish the importance of the micropylar plug for germination. The breaking of dormancy requires storage for the length of the dry season in the tropical deciduous forest, the habitat of these plants.

Annona macroprophyllata conocida comúnmente como papausa o ilama es una especie frutícola de importancia económica, consumida en el sur de México y en América central; es considerada por su sabor, el fruto más fino de las anonáceas. Esta especie presenta problemas de propagación debido al periodo de latencia de hasta seis meses que tienen sus semillas, el cual no ha sido completamente explicado. Entre los factores que impactan la germinación de las semillas, están la posición de estas dentro del fruto y la presencia de un tapón micropilar leñoso. El presente trabajo investigó la importancia de estos factores relacionados a la germinación y latencia de esta especie. Para este estudio, se estructuró un diseño completamente al azar usando 169 frutos de Annona macroprophyllata de la variedad blanca. Los frutos fueron divididos en tres secciones transversalmente (basal, medio y apical), las semillas de cada sección fueron caracterizadas en tamaño y peso y por nueve meses fueron almacenadas y evaluadas con relación a su viabilidad y capacidad germinativa, considerando al mismo tiempo la presencia o ausencia del tapón micropilar. Los resultados no correlacionan la posición de las semillas en el fruto, ni el tapón micropilar con el rompimiento de la latencia, pero revelan la importancia del tapón micropilar para la germinación. El rompimiento de la latencia de esta especie depende del tiempo de almacenamiento, el cual está completamente asociado a la temporada de sequía de la selva baja caducifolia, donde esta especie habita.

Estudos químicos em plantas da família Annonaceae se intensificaram nas últimas décadas, devido à descoberta de moléculas de anonáceas com potencial medicinal (por exemplo, alcalóides benzilisoquinolina e acetogeninas). Aproximadamente 500 alcaloides têm sido identificados em 138 espécies de Annonaceae em 43 gêneros. Além disso, até 2004, 593 acetogeninas de annonaceas (ACGs) foram identificados, a partir de 51 espécies em 13 gêneros. Isto sugere que as plantas desta família atribuem recursos importantes para a biossíntese destes compostos. Apesar da diversidade destas moléculas, suas funções biológicas, incluindo as suas funções fisiológicas e/ou ecológicas, não são bem compreendidas. Neste estudo, foram fornecidos novos dados que descrevem a variedade e a distribuição de certos alcaloides e ACGs em annonaceas em fases distintas de desenvolvimento. As possíveis relações entre alguns destes compostos e as alterações climáticas sazonais ocorrendo no habitat são também discutidas. Estes dados vão melhorar a nossa compreensão do metabolismo secundário dessas moléculas farmacologicamente importantes e seus padrões de expressão durante o desenvolvimento, o que ajudará a determinar as condições ideais de crescimento e épocas de colheita para a sua produção.

Chemical studies of the plant family Annonaceae have intensified in the last several decades due to the discovery of annonaceous molecules with medicinal potential (e.g., benzylisoquinoline alkaloids and acetogenins). Approximately 500 alkaloids have been identified in 138 Annonaceae species in 43 genera. In addition, until 2004, 593 annonaceous acetogenins (ACGs) had been identified, from 51 species in 13 genera.This suggests that plants from this family allocate important resources to the biosynthesis of these compounds. Despite the diversity of these molecules, their biological roles, including their physiological and/or ecological functions, are not well understood. In this study, it was provided new data describing the variety and distribution of certain alkaloids and ACGs in annonaceous plants in distinct stages of development. The potential relationships among some of these compounds and the seasonally climatic changes occurring in the plant habitat are also discussed. These data will improve our understanding of the secondary metabolism of these pharmacologically important molecules and their expression patterns during development, which will help to determine the optimal growth conditions and harvest times for their production.

Los alcaloides bencilisoquinolínicos (ABI) son metabolitos especializados con una distribución filogenética antigua pero conservada todavía en clados modernos. Varios de ellos, como la morfina, sanguinerina y berberina tienen importancia en la medicina moderna. En esta revisión se analizan los aspectos más sobresalientes del estado actual de la biosíntesis de ABI. Se han realizado estudios que han permitido conocer la biosíntesis de 22 de estos metabolitos nitrogenados. En su formación participan 43 enzimas agrupadas en oxido-reductasas, transferasas y liasas, que en algunos casos representan ejemplos atípicos de la forma en la que se originó la diversificación del metabolismo secundario, entre ellos proteínas citocromo P450 (CYP450) con actividades catalíticas para la ruta de los ABI, o la enzima norcoclaurina sintasa (NCS) que esta emparentada con proteínas alergénicas de defensa. Así mismo, hay avances genéticos en los que se ha podido caracterizar 30 enzimas, permitiendo conocer procesos de regulación. Otro aspecto interesante es la compartimentación de los sitios de biosíntesis y acumulación de ABI ya que en varios casos están separados espacialmente y en distintas especies o en la misma pueden participar varios tipos de células. Ello ha sugerido el transporte intra e intercelular de los alcaloides, los precursores y de las enzimas, se ha documentado el transporte de berberina entre el citoplasma y las vacuolas del almacenamiento. El panorama de la biosíntesis de ABI se ha construido con los estudios de ejemplares de importancia farmacológica.

The benzylisoquinoline alkaloids (BIA) are specialized metabolites with an ancient phylogenetic distribution, but still preserved in modern clades. Some of them, such as morphine, sanguinerine or berberine, are important for modern medicine. This review discusses the highlights of the current state of the biosynthesis of BIA. There have been studies that show the biosynthesis of 22 of these nitrogenous metabolites. In their formation there are 43 enzymes grouped into oxidoreductases, transferases and lyases, which in some cases represent atypical examples of the manner in which the secondary metabolism diversification was originated. Two of these examples are the cytochrome proteins P450 (P450), with catalytic activities for ABI route, or the norcoclaurine synthase enzyme (NCS), which share substantial identity with defense allergenic proteins. Likewise, there are genetic advances that have produced the characterization of 30 enzymes, allowing knowledge of regulatory processes. Another interesting aspect is the compartmentation of the biosynthesis sites and accumulation of BIA, since in several cases they are spatially separated and in different species, or in the same species several types of cells may be involved. This has suggested intra and intercellular transport of alkaloids, precursors and enzymes, and it has been documented berberine transport between the cytoplasm and the vacuoles of storage. The picture for the biosynthesis of BIA has been constructed with exemplary studies of alkaloids with pharmacological importance.

Os alcalóides benzilisoquinolinas (ABI) são metabólitos especializados com uma distribuição filogenética antiga, mas ainda preservada em clados modernos. Vários deles, como a morfina, sanguinarina e berberina são importantes na medicina moderna. Neste artigo, se analisam os aspectos mais destacados do estado atual da biossíntese de ABI; há estudos que tem permitido conhecer a biossíntese de 22 desses metabólitos nitrogenados. Na sua síntese participam 43 enzimas agrupadas em oxidoreductases, transferases, liases e, em alguns casos, representam exemplos atípicos da forma pela qual se originou a diversificação do metabolismo secundário, incluindo as proteínas do citocromo P450 (CYP450), com atividades catalíticas para a rota dos ABI, ou a enzima norcoclaurina sintase (NCS), que está relacionada com proteínas alergênicas de defesa. Da mesma forma, há avanços genéticos na caracterização de 30 enzimas, permitindo conhecer processos de regulação. Outro aspecto interessante é a compartimentalização dos sítios de biossíntese e acumulação de ABI uma vez que em muitos casos estão separados espacialmente e em diferentes espécies, ou na mesma podem participar vários tipos de células. Isto há sugerido o transporte intra e intercelular de alcalóides, precursores das enzimas; tem sido documentado o transporte de berberina entre o citoplasma e os vacúolos de armazenamento. A perspectiva na biossíntese de ABI foi construída com os estudos de exemplares de importância farmacológica.