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The benzylisoquinoline alkaloids of Annona muricata have been isolated, but their physiological or ecological role is unknown. The objective was to explore whether these secondary metabolites are involved in defense against phytopathogenic fungi. To do this, the alkaloidal response of 6-leaf seedlings of A. muricata was analyzed, previously inoculated with Colletotrichum gloeosporioides and Rhizopus stolonifer. Before and after inoculation, alkaloidal extracts of roots, stems, and leaves were obtained, and the antifungal activity was evaluated in vitro. The alkaloids anonaine, reticuline, nornuciferine, assimilobine, and coreximine were identified. C. gloeosporioides caused variable increases in the production of anonaine, reticuline and nornuciferine (10–1200%), while R. stolonifer only stimulated the increase of nornuciferin and anonaine (10%) in the stems and leaves. The alkaloidal extracts of inoculated seedlings increased the antifungal activity, both against the pathogen elicitor and against the second target pathogen. These findings suggest that the alkaloids participate in the antifungal defense mechanism.

The benzylisoquinoline alkaloids of

Made available in DSpace on 2019-10-06T16:25:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2019-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) The Annonaceae family presents alkaloids with ecological functions and pharmacological interest. This is the first study to evaluate if the application of plant growth regulators to seeds of this family alters the production of alkaloids over time from germination. This study was carried out in four sequential stages of Annona cacans Warm. development from the resumption of embryo during seed imbibition with the application of plant regulator GA4+7 + 6-Benzyladenine. The concentration of total alkaloids was quantified using the oxoaporphine alkaloid liriodenine as reference standard. In addition, the liriodenine concentration was measured and the profile evaluated by ultra-high-performance liquid chromatography (UHPLC). Results have shown that alkaloids are present in all phases and in all tissues, at higher concentrations in roots (up to 100 times). The proportion of total alkaloids and liriodenine was modified in response to the application of plant regulators. Roots doubled the content of alkaloids and liriodenine. In cotyledonary leaves, the amount of total alkaloids decreased; however, liriodenine remained unchanged. Our results have shown that the use of plant growth regulators based on gibberellins and cytokinins modified the production of alkaloids in tissues in a specific way. Department of Botany São Paulo State University (UNESP) Institute of Biosciences Botucatu, Rua Prof. Dr. Antonio Celso Wagner Zanin, 250 Laboratorio de Fisiología y Química Vegetal Instituto de Ciencias Biológicas Universidad de Ciencias y Artes de Chiapas (UNICACH), Libramiento Norte Poniente 1150, Col. Lajas Maciel Center for Studies and Research of Medicinal Plants (NEPLAME) Federal University of Vale do São Francisco Departament of Chemistry Federal University of Amazonas Federal Institute of Paraná (IFPR), Av. das Pombas, 2020 Department of Botany São Paulo State University (UNESP) Institute of Biosciences Botucatu, Rua Prof. Dr. Antonio Celso Wagner Zanin, 250

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. Interacción de Bephratelloides cubensis (Hymenoptera: Eurytomidae) con frutos de Annona macroprophyllata en huertos de Chiapas, México. 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.

Made available in DSpace on 2021-06-25T11:07:34Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-01-01. Added 1 bitstream(s) on 2021-07-15T14:36:01Z : No. of bitstreams: 1 S0100-29452020000602002.pdf: 4708987 bytes, checksum: 25bf646d264e1feec456206002a406a7 (MD5) Studies on the effect of the applying of phytoregulators to overcome seed dormancy have been carried out in Annonaceae species, thus the endogenous relationship between abscisic acid and gibberellins after the application of phytoregulators needs to be known in these species. Considering the importance of hormonal balance in dormancy mechanisms, the aim of this research was to elucidate how endogenous ABA and GA concentrations change after the application of phytoregulators in seeds and how these changes affect reserve degradation during germination of Annona macroprophyllata and A. purpurea seeds. Seeds were submitted to three conditions:-no soaking, soaking-in-water and soaking-in-GA4+7 +benzyladenine. ABA, GA, lipids, proteins, and total soluble sugars were quantified at 0, 2, 5, 10, and 15 days after the beginning of treatments. The application of phytoregulators led change of the hormonal balance inducing increase in endogenous GA and reduction in ABA levels since seed soaking. During imbibition, the degradation of seed reserves (breaks proteins first, and then soluble sugars and finally lipids) was observed and complete germination was obtained after 10 days, with primary root emission. In both species, dormancy can be broken soon after seed dispersal when the balance between GA and ABA is endogenously changed (by exogenous application of phytoregulators), leading to metabolic reserve degradation and germination. Plant Biology BBVPZ Department Institute of Biosciences São Paulo State University (Unesp) Instituto de Ciencias Biológicas Universidad de Ciencias y Artes de Chiapas (UNICACH) Plant Biology BBVPZ Department Institute of Biosciences São Paulo State University (Unesp)

Several species of Solanum produce secondary metabolites with antimicrobial activity. In the present study, the inhibitory activity of Solanum chrysotrichum, S. erianthum, S. torvum and S. rostratum against phytopathogenic Curvularia lunata was determined. Methanol extracts from roots, stems, leaves and fruits were evaluated by the method of mycelial inhibition on agar and the minimum inhibitory concentration (MIC) was determined on a liquid medium. To increase the antimicrobial activity, the combined activity of the most active extracts for each phytopathogen was also determined (a combination of intra and interspecies extracts). The results showed that 12 of the 16 methanolic extracts of Solanum species had antifungal effects against C. lunata. The extracts of S. rostratum and S. erianthum developed the highest activity (~80% inhibition and 28.4 MIC μg . ml–1), even, equal to or greater than, the reference fungicide. The mixture of the active extracts of S. chrysotrichum and S. torvum increased their activity. Various extracts affected the macro and microscopic morphology and most of them reduced the number of conidia of the fungus. This resulted in the capacity to control the vegetative growth and reproduction of C. lunata, the causal fungus of corn leaf spot disease.

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.

In plants, the presence and distribution of specialized metabolites during the early stages of development are not documented enough, even though their biosynthesis is one of the most important strategies for survival. In this study, five alkaloids and four acetogenins were detected in Annona muricata L. during early development seedling, including three phases of root emergence and three of seedling formation. Hexane and alkaloid extracts were obtained from each organ, which were analyzed in a gas-mass chromatograph and in a high-performance liquid chromatograph coupled with a photodiode array UV detector (HPLC-DAD). This research shows the presence of the acetogenins cis-uvarimicin IV, mosinone, muricina B, and cis-annonacin-10-one, as well as of the alkaloids reticuline, coreximine, anonaine, asimilobine, and nornuciferine, both groups with a variable organ-specific distribution, related with the formation of organs and tissues.

Made available in DSpace on 2021-06-25T10:46:22Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-11-28. Added 1 bitstream(s) on 2021-07-15T15:23:18Z : No. of bitstreams: 1 S1678-91992020000100337.pdf: 7061245 bytes, checksum: f1175c6ed210c2f03ef23c270bd20f59 (MD5) 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 noninfectious 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. Department of Tropical Disease and Imaging Diagnosis Botucatu Medical School São Paulo State University (UNESP) Universidad de Ciencias y Artes de Chiapas Institute of Biosciences of Botucatu São Paulo State University (UNESP) Faculdade de Ciências da Saúde Universidade do Oeste Paulista (Unoeste) Department of Tropical Disease and Imaging Diagnosis Botucatu Medical School São Paulo State University (UNESP) Institute of Biosciences of Botucatu São Paulo State University (UNESP)

Background and Aims: The phytochemistry study of Annonaceae has intensified in the last decades due to the discovery of secondary metabolites with antimicrobial activities. The inhibitory activities of extracts and compounds from these species for phytopathogens are less known. The antifungal activity of Sapranthus microcarpus was determined in vitro against six fungal pathogens of important crops. Methods: Hexane, methanol, and alkaloid extracts of the leaves and root and stem bark of S. microcarpus were evaluated to determine whether they could inhibit the mycelial growth of the phytopathogens. The combined activity of the most active extracts for each phytopathogen was also determined. The alkaloid liriodenine was isolated and identified as an antifungal principle. Key results: All extracts inhibited the growth of the six phytopathogens to some extent. The root bark alkaloid extract showed the highest activity. Fusarium oxysporum f. sp. lycopersici and Curvularia lunata were the most sensitive phytopathogens. Liriodenine was active against all the plant pathogens (minimum inhibitory concentrations of 125-500 nmol ml -1 ). Conclusions: Sapranthus microcarpus is a native natural resource with great phytochemical potential.

Annona purpurea grows in the areas of low elevation in deciduous forests of Mexico, those areas have marked rainy and dry seasons. This species produces more than 30 bioactive alkaloids that could have potential in the control of phytopathogens. This research provides data on the variation of the content and number of alkaloids during an annual cycle and the associated inhibitory potential of the compounds against three phytopathogenic fungi. For one year, alkaloidal extracts of stems and leaves were obtained every two months. The extract profiles were determined by gas chromatography with tandem mass spectrometry and their antifungal activity was examined in vitro. The alkaloids, annomontine and oxopurpureine, obtained from the roots and leaves, respectively, were also evaluated individually. The yields, profiles and activities of the extracts, as well as the abundance of annomontine and oxopurpureine in the extracts, were contrasted with the seasonality and phenological phases of the plant. The data indicate that the alkaloid content was higher at the height of the dry season. High yields also occurred during flowering. The strongest inhibitory effect was obtained from the root extracts during the last month of dry season. This finding seems to be explained by the higher chemodiversity of alkaloids in extracts from this season. Annomontine and oxopurpureine inhibited all three phytopathogens; however, they were not solely responsible for the activity of A. purpurea.

The seeds of Annona macroprophyllata Donn. Sm. contain idioblasts with toxic acetogenins, including laherradurin and rolliniastatin-2, in relatively high proportions. Both metabolites are cataloged as potent insecticides for several species, even so, the wasp Bephratelloides cubensis Ashmead fulfills almost its entire life cycle inside the seeds of this and other annonaceous species, to such a degree, that they constitute a strong selection pressure. In order to document the chemical relationship between the two species, it is reported for the first time in this paper the presence of idioblasts and acetogenins during the ontogenic development of the seeds of A. macroprophyllata, and contrasted with the development of B. cubensis. The results indicate that idioblasts with laherradurin and rolliniastatin-2 acetogenins are formed in the middle stages of the endospermic development, also that both acetogenins are biosynthesized simultaneously, and that their proportion is dependent on the degree of development. The acetogenins are present in high amounts that suppose a sufficient toxic barrier and, in this case, laherradurin is the most abundant (> 1000 µg g dry weight−1). The wasp B. cubensis only emerges from the seeds to copulate and returns for oviposition; its larval phase coincides with the appearance of acetogenins, so it feeds on the acetogenic endosperm. The absence of acetogenins in the tissues and excreta of the insect supposes a metabolization of the molecules, which would explain the tolerance to its toxicity.

Annonaceous acetogenins (ACGs) are molecules with carbon numbers C35-C37, usually with tetrahydrofuran and tetrahydropyran rings and one terminal γ-lactone (usually α,β-unsaturated), in a large aliphatic chain that is varyingly hydroxylated, acetoxylated or ketonized. ACGs have ecological functions as insecticides and are pharmacologically promising due to their cytotoxic and antitumoral properties. They are found in the seeds, leaves, roots, flowers and fruits of annonaceous plants and can be detected during isolation via thin-layer chromatography using Kedde's reagent, which reacts with the unsaturated lactone. This chapter describes the location in situ of ACGs in fresh sections of annonaceous seeds using Kedde's reagent.The acetogenins are located in the idioblasts, in the endosperm and in the embryonic axis during differentiation. This method can aid in the detection of ACGs with a terminal unsaturated γ-lactone in organs and tissues.

Annonaceous acetogenins (ACGs) are molecules with carbon numbers C35-C37, usually with tetrahydrofuran and tetrahydropyran rings and one terminal γ-lactone (usually α,β-unsaturated), in a large aliphatic chain that is varyingly hydroxylated, acetoxylated or ketonized. ACGs have ecological functions as insecticides and are pharmacologically promising due to their cytotoxic and antitumoral properties. They are found in the seeds, leaves, roots, flowers and fruits of annonaceous plants and can be detected during isolation via thin-layer chromatography using Kedde's reagent, which reacts with the unsaturated lactone. This chapter describes the location in situ of ACGs in fresh sections of annonaceous seeds using Kedde's reagent.The acetogenins are located in the idioblasts, in the endosperm and in the embryonic axis during differentiation. This method can aid in the detection of ACGs with a terminal unsaturated γ-lactone in organs and tissues.

Currently, the Annonaceae family is characterised by the production of acetogenins (ACGs), and also by the biosynthesis of alkaloids, primarily benzylisoquinolines derived from tyrosine. The objective of this study was to confirm the presence of alkaloids and acetogenins in the idioblasts of the endosperm and the embryonic axis of A. macroprophyllata seeds in germination. The Dragendorff, Dittmar, Ellram, and Lugol reagents were used to test for alkaloids, and Kedde's reagent was used to determine the presence of acetogenins in fresh sections of the endosperm and embryonic axis of seeds after twelve days of germination. A positive reaction was observed for all the reagents, and the presence of alkaloids and acetogenins was confirmed in the idioblasts of the endosperm and those involved in the differentiation of the embryonic axis of the developing seedling. We concluded that the idioblasts store both metabolites, acetogenins and alkaloids. Beginning at differentiation, the idioblasts of the embryonic axis simultaneously biosynthesise acetogenins and alkaloids that are characteristic of the species during the development of the seedling. The method used here can be applied to histochemically confirm the presence of acetogenins and alkaloids in tissues and structures of the plant in different stages of its life cycle.

Early development is a critical stage in a plant’s life, as the plant must establish itself in the ecosystem during this period. The secondary metabolites (SM) during this phase is a strategy that contributes to the survival of plant species. Through a review of the literature, a number of reports were found that investigated the presence of SM during germination and early plant development (phases 0 and 1 according to the Zadoks and BBCH scales). A total of 250 reports were found that investigated 99 species and nearly 200 SM that accumulate during this period of the plant life cycle. A large portion of the SM are biosynthesised de novo, whereas the remainder are derived in part or in total from the mother plant. In many cases, the resources for biosynthesis are supplied only by the reserve material of the endosperm or cotyledons, which allows for independent photosynthesis. The presence of SM at these stages confers characteristics of more advanced stages, such as tissue-specific distribution, spatio-temporal regulation, and the individual regulation of all of the biosynthesised SM. The amount and diversity of SM are not universally related to the progress of plant development, but it is a widespread phenomenon. The early production of SM has ecological implications that involve defence mechanisms, relationships with microorganisms, and the role of these compounds as nitrogen reserves. This review contributes to the systematisation of studies on SM in the early stages of development.

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 wasoriginated. 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.Key words: specialized metabolism, secondary metabolism, cellular transport, cell compartment, tissue-specific regulation

The effects of annomontine, a pyrimidine- β-carboline alkaloid isolated from the root of ANNONA PURPUREA, on anxiety was studied in mice using the elevated plus-maze. The behavioral effects of this alkaloid on the pentobarbital-induced hypnosis, the locomotor activity in an open field, and the motor coordination in the rotarod test were also evaluated. The intraperitoneal injection of annomontine (1-30 mg/kg) increased in a dose-dependent way the number of visits to and the time spent in the open arms of the elevated plus-maze in comparison to the control animals. Such effects were blocked by the prior application of flumazenil (3 mg/kg; i. p.), a specific antagonist for the binding of benzodiazepines on the GABA (A) receptor. Under the same experimental conditions annomontine failed to affect the behavior of the animals in the pentobarbital-induced hypnosis test and had no effects on locomotion and motor coordination. These results suggest that annomontine possesses anxiolytic-like effects which may be mediated at the level of the benzodiazepine binding site on the GABA (A) receptor.

Rolliniastatin-2 is an acetogenin with elevated cytotoxic activity whose effect on bacteria has not yet been analysed and was isolated by CC from the hexanic extract of Annona diversifolia Saff (Annonaceae) seeds. In this study, we evaluated the susceptibility of Escherichia coli, Pseudomonas aeruginosa and Salmonella typhi as well as spheroplasts of these bacteria to the compound, determining percent inhibition by turbidimetry. The complete bacteria were susceptible to rolliniastatin-2 (28-40% at 2 mg mL(-1)), while spheroplasts showed an increased susceptibility (55-58%, at 1 ng mL(-1)). The most susceptible bacterium was P. aeruginosa, while the spheroplasts of E. coli and S. typhi were most sensitive to the compound's effects.

Aporphine alkaloids are secondary metabolites that are obtained in low levels from species of the Annonaceae family. Nitrogen addition may increase the alkaloid content in plants. However, previous studies published did not consider that nitrogen could change the alkaloid content throughout the day. We conducted this short-term study to determine the effects of nitrogen applied throughout the diurnal period on the aporphine alkaloids via measurements conducted on the roots, stems and leaves of Annona diversifolia seedlings. The 60-day-old seedlings were cultured with the addition of three levels of nitrogen (0, 30 and 60 mM), and alkaloid extracts were analysed using high-performance liquid chromatography. The highest total alkaloid content was measured in the treatment with moderate nitrogen supply. Further, the levels of aporphine alkaloids changed significantly in the first few hours of the diurnal period. We conclude that aporphine alkaloid content increased with moderate nitrogen supply and exhibited diurnal variation.

Some Annonaceae seeds are known to exhibit dormancy mechanisms ranging from possible seed coat impermeability to physiological dormancy. Thus, the aim of this study was to evaluate the effects of gibberellin (GA) GA3 and GA4+7 + benzyladenine (GA4+7 + BA) application in seeds of Annona macroprophyllata Donn. Sm (papausa) and Annona purpurea Moc. & Sessé ex Dunal (chincuya). The experiment was performed by the application of GA3 and GA4+7 + BA on seeds in concentrations of 0, 200, 400, 500, 600, 800 and 1000 mg L-1. The regulators broke the dormancy of both species. However, application of the GA4+7 + BA mixture had more significant results, with greater increases in germination in A. macroprophyllata than in A. purpurea. Treatments that promoted the highest germinations were GA4+7 + BA at a concentration of 200 mg L-1 for A. macroprophyllata (77%) and 200 mg L-1 of GA4+7 + BA and 500 mg L-1 of GA3 for A. purpurea (30% and 29%, respectively). Rate index, mean time and frequency of germination were distinct for both species and both treatments. Although both GA3 and GA4+7 + BA promote germination, the GA4+7 + BA mixture was more effective than GA3 to overcoming seed dormancy of both species, A. purpurea has a harder dormancy than A. macroprophyllata RESUMO Sementes de Annonaceae são conhecidas por possuírem mecanismos de dormência que variam de impermeabilidade de tegumento até à dormência fisiológica. Desta forma, o objetivo deste estudo foi avaliar os efeitos da giberelina GA3 e de GA4+7+ a citocinina benziladenina (GA+BA) na superação da dormência de sementes de Annona macroprophyllata Donn. Sm (papausa) e Annona purpurea Moc. & Sessé ex Dunal (chincuya). O experimento foi realizado com a aplicação de GA3 e GA4+7 + BA nas concentrações de 0; 200, 400, 500, 600, 800 e 1.000 mg L-1. O uso dos reguladores resultou na quebra da dormência em ambas as espécies. Contudo, a aplicação da mistura GA4+7 + BA resultou em maiores incrementos na germinação de A. macroprophyllata do que em A. purpurea. Os tratamentos que promoveram as maiores porcentagens de germinação foram 200 mg L-1 de GA4+7 + BA para A. macroprophyllata (77%) e 200 mg L-1 de GA4+7 + BA e 500 mg L-1 de GA3 para A. purpurea (30% e 29%, respectivamente). O índice de velocidade, o tempo médio e a frequência de germinação foram diferentes para ambas as espécies e com ambos os reguladores. Conclui-se que, apesar de GA3 e GA4+7 + BA promoverem a germinação, a mistura GA4+7 + BA foi mais efetiva que GA3 para quebrar a dormencia de ambas as espécies. Além disso, sementes de A. purpurea apresentam dormência mais difícil de ser superada que A. macroprophyllata.

Acetogenins (ACGs) are bioactive compounds with cytotoxic properties in different cell lines. They are antitumoural, antiparasitic, antimalarial, insecticidal, antimicrobial, antifungal and antibacterial. These secondary metabolites function in plant defence and are found in specific organelles and specific cells, thereby preventing toxicity to the plant itself and permitting site-specific defence. The aim of this work was to histochemically determine the in situ localisation of ACGs in the endosperm of Annona macroprophyllata seeds using Kedde’s reagent. Additionally, the colocalisation of ACGs with other storage molecules was analysed. The seeds were analysed after 6 and 10 days of imbibition, when 1 or 2 cm of the radicle had emerged and metabolism was fully established. The seeds were then transversally cut in half at the midline and processed using different histological and histochemical techniques. Positive reactions with Kedde’s reagent were only observed in fresh, unfixed sections that were preserved in water, and staining was found only in the large cells (the idioblasts) at the periphery of the endosperm. The ACGs’ positive reaction with Sudan III corroborated their lipid nature. Paraffin sections stained with Naphthol Blue Black showed reactions in the endosperm parenchyma cells and stained the proteoplasts blue, indicating that they might correspond to storage sites for albumin-like proteins. Lugol’s iodine, which is similar in chemical composition to Wagner’s reagent, caused a golden brown reaction product in the cytoplasm of the idioblasts, which may indicate the presence of alkaloids. Based on these results, we propose that Kedde’s reagent is an appropriate histochemical stain for detecting ACGs in situ in idioblasts and that idioblasts store ACGs and probably alkaloids. ACGs that are located in idioblasts found in restricted, peripheral areas of the endosperm could serve as a barrier that protects the seeds against insects and pathogen attack.

Made available in DSpace on 2014-10-01T13:08:41Z (GMT). No. of bitstreams: 0 Previous issue date: 2014Bitstream added on 2014-10-01T14:03:54Z : No. of bitstreams: 1 S0100-29452014000500034.pdf: 496147 bytes, checksum: 8708146290c98cf387e11c7889ce1398 (MD5) As sementes de espécies da família Annonaceae são conhecidas por apresentarem mecanismos de dormência, cujos relatos vão desde a impermeabilidade do tegumento até a dormência fisiológica. Neste sentido, o objetivo do presente trabalho foi avaliar a impermeabilidade à água das sementes de Annona diversifolia Saff e Annona purpurea Moc & Sessé ex Dunal a partir do estudo da curva de aquisição de água. Para a realização do estudo, sementes foram colocadas sob imersão em água destilada, sendo utilizadas 4 repetições de 25 sementes de cada espécie, as quais foram pesadas durante as 480 horas em que ficaram imersas. Para determinar o local de aquisição de água, sementes de Annona diversifolia foram vedadas em diferentes locais com parafina. Com base nos resultados, pode-se afirmar que sementes de ambas as espécies apresentam Fase I e II da germinação, o que indica que as sementes não são impermeáveis; contudo, a Fase III não é alcançada. As sementes de Annona diversifolia completam a Fase I com 50 horas, e as de Annona purpurea, com 70 horas do início da embebição, o que demonstra que, mesmo de maneira lenta, a água é adquirida. Annonaceae seeds are known by presenting dormancy mechanisms, whose reports ranging from coating impermeable to the physiological dormancy. By this way, the present study aimed to evaluate water uptake in Annona diversifolia Saff and Annona purpurea Moc & Sessé ex Dunal seeds. For this study, seeds were placed under immersion in distilled water, and used four replicates of 25 seeds of each species, which were weighed during the 480 hours that were immersed. To determine the place of purchase of water, Annona diversifolia seeds were sealed with paraffin at different locations. Based on the results, seeds from both species reached the phases I and II of water uptake, which indicates they are not hard; however, germination (Phase III) was not reached. Annona diversifolia seeds completed Phase I with, 50h and Annona purpurea with 70h from imbibitions begin, which shows that even slowly, water is acquire. UNESP IBB Depto Botânica Universidad de Ciencias y Artes de Chiapas Faculdade de Ciências Biológicas UNICACH Faculdade de Ciências Biológicas UNESP IBB Depto Botânica

Background: For many years Chiapas, Mexico's poorest state, has had the highest rate of child mortality due to intestinal infections of unknown etiology in the country. To begin identifying the infectious agents, our work determined the prevalence of intestinal parasites as well as malnutrition in children from Chiapas's three most impoverished municipalities: Pantepec, Chanal, and Larrainzar. Methodology: In this cross-sectional study, conducted between January and November 2009, we assessed the prevalence of intestinal parasites by means of coproparasitological analysis in children 40% of children represented varying degrees of malnutrition and a marked constitutional delay in growth. A very high prevalence of stunting was also recorded in children from Chanal and Larrainzar (70% and 55 %, respectively). An association between infection with intestinal parasites and malnutrition was observed in Pantepec. Preschool-age children were more likely to be infected with intestinal parasites. Conclusion: Our results indicate the urgent need for interventions in order to 1) improve the nutritional status of children and 2) reduce infection rates of enteric parasites.

Annona lutescens Saff. (Annonaceae) grows as a native tree in Chiapas, Mexico in Tropical Dry Forest habitat. Like most Annonaceae, it biosynthesizes benzylisoquinoline alkaloids, mostly liriodenine. To determine the influence of seasonal changes in the accumulation of liriodenine, the monthly variation of liriodenine content in roots, stems and leaves of mature and young trees was observed. These parts of young and mature A. lutescens trees were collected monthly over a 1 year period and the alkaloids were extracted; the liriodenine was quantified by high-resolution liquid chromatography. The phenological stages of the species were also assessed (leaf development, flowering and fruiting) using the Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scale. The analysis of both young and mature trees showed a significant increase in the liriodenine concentration occurs within roots during the dry season, which coincides with leaf fall. A significant decrease also occurred at the beginning of the rainy season (the period of leaf growth); the liriodenine content for the next rainy season did not reach the levels of the previous dry season. The climatic variation induced phenological and physiological changes in this species.

Annonaceae (custard apple family) extracts have demonstrated their insecticidal potential against a variety of insect pests. This study is aimed to determine the activity of ethanolic and aqueous extracts of the stems and leaves of Annona muricata L., A. diversifolia Saff. and A. lutescens Saff. against larvae of Anastrepha ludens (Mexican fruit fly). Ethanolic extracts were obtained by Soxhlet extraction, and aqueous extracts were obtained by boiling. Larvicidal activity against third in star A. ludens larvae was determined after 24 and 72h of larval exposure to the extracts at concentrations of 100, 1000 and 2000ug x ml-1. Additionally, toxicity was quantified by the Artemia salina (brine shrimp) assay. The results indicate that extracts of the three aforementioned Annona species show time-dependent larvicidal activity against A. ludens, with variable mortality rates at 72h of exposure, as follows: A. lutescens 87- 94%, A. diversifolia 70-90% and A. muricata 63-74%

Annonaceae aporphine alkaloids, of which liriodenine is the most abundant, have not been extensively studied from a biological standpoint. The goal of this study was to investigate the role of liriodenine in antimicrobial defense during early developmental stages in Annona diversifolia. The fungi Rhizopus stolonifer and Aspergillus glaucus, which are responsible for seed deterioration, were isolated during imbibition, and their antifungal activity was determined by diffusion, macrodilution, and metabolic inhibition assays using purified liriodenine and alkaloid extracts obtained from embryos, radicles, and roots at early developmental stages. The presence of liriodenine in extracts was quantified by high-performance liquid chromatography. Purified liriodenine and alkaloidal extracts inhibited both fungi, and there was a positive relationship between extract activity and amount of liriodenine contained therein. The quantity of liriodenine present in extracts suggests its importance in controlling other phytopathogens.

Plants of the Annonaceae family produce a series of alkaloids, including liriodenine oxoaporphine. Its distribution in these primitive angiosperms suggests that it plays an important role, but very little is known about which plant organs it accumulates in, or in which developmental stages it is synthesised. Accordingly, liriodenine production was studied during the early stages of germination and seedling development in Annona diversifolia Saff. Liriodenine samples were obtained from the roots and were characterised on the basis of spectroscopic data. Quantification was done by HPLC in the organs and tissues of newly collected seeds, seeds following 1-, 2-, 5- and 10-day imbibitions, upon emergence of the radicle and at the seedling stage. According to our results, liriodenine could not have originated from the parent plant, nor during embryogenesis because it appears for the first time in the endosperm approximately 5 days after the start of imbibition. Therefore, its synthesis does not depend directly on photosynthesis. During the seedling stage it is found in the root and the stem but it is absent from the cotyledonary leaves and the first true leaves. Liriodenine biosynthesis begins during the early stages of development in the endosperm and seed radicles.

This study evaluated possible CNS effects of a hexane extract of leaves from Rollinia mucosa (Jacq.) Baill. (Annonaceae). This plant extract induced anxiolytic-like actions similar to those induced by diazepam in the avoidance exploratory behavior paradigm. Its significant activity was shown at doses from 1.62 to 6.25 mg/kg. It also enhanced pentobarbital-induced hypnosis time, and at high doses produced motor coordination impairment. The benzodiazepine (BDZ) receptor binding, evaluated by in vitro autoradiography following a single administration of R. mucosa, revealed that this plant extract reduced BDZ binding in the hippocampus (29%), amygdala (26%), and temporal cortex of mice (36%). In conclusion, the present findings support the proposal that R. mucosa may induce central nervous system (CNS) depressant effects, presumably through an interaction with the GABA/benzodiazepine receptor complex.

Acetogenins from Annonaceae (ACG) are potent inhibitors of the mitochondrial complex I, they present cytotoxic activity on neoplasic lines, including those with multiresistance to drugs. In vivo antitumor activities of some ACG have been reported; however, no information is available regarding the relationship between their cytotoxic activity in cell cultures and their antiproliferative action in vivo. Laherradurin and cherimolin-2 acetogenins were isolated from Annona diversifolia seeds, and their inhibitory potential was analysed in vitro on HeLa and SW-480 cells. Doses containing 1, 10, 100 and 500 times the IC50 obtained from the proliferation assays and multiplied by the weight of the animal, were injected once daily into athymic mice bearing these cancer cell lines; their effect upon tumor growth was measured over a period of 20 days. Laherradurin was more active than cherimolin-2, and it showed in in vitro proliferation assays a similar IC50 in both neoplasic lines. In athymic mice, laherradurin administered in 10x, 100x and 500x doses, reduced the size of HeLa tumors, and with 100x and 500x doses, affected the SW-480 tumor development. These doses were similar to results found with the control drug doxorubicin (p < or = 0.05). On the other hand, cherimolin-2 had an effect on HeLa tumors cells at 100x and 500x doses (p < or = 0.05).

Background Mushrooms generate strong and contrasting feelings ranging from extreme aversion to intense liking. To categorize these attitudes, Wasson and Wasson coined the dichotomic terms “mycophilia” and “mycophobia” in 1957. In Mesoamerica these categories have been associated to ecological regions. Highland peoples are viewed as mycophiles, whereas lowland inhabitants are considered mycophobes. However, this division is based on little empirical evidence and few indicators. This study questioned whether mycophilia and mycophobia are indeed related to ecological regions through the evaluation of 19 indicators tested in the highlands and lowlands of Chiapas, Mexico. Methods The heterogeneity of attitudes toward mushrooms was explored in terms of ecological region and sociocultural variables. Information was obtained through structured interviews in 10 communities in Los Altos de Chiapas (highlands) and the Selva Lacandona (lowlands). We analyzed indicators separately through χ2 tests and multivariate techniques. The Mycophilia-Mycophobia Index was also used in the analysis. To assess which factors better explain the distribution of attitudes, we built 11 models using the Beta probability-density function and compared them with the Akaike Information Criterion. Results Most people had positive attitudes in both ecological regions. The classification and ordination analyses found two large groups comprising both highland and lowland towns. Contrary to expectation if mycophilia and mycophobia were mutually exclusive, all the fitted probability distributions were bell-shaped; indicating these attitudes behave as a continuous variable. The model best supported by data included occupation and ethnicity. Indigenous peasants had the highest degree of mycophilia. Discussion Results suggest the studied populations tend to be mycophilic and that their attitudes are not dichotomic, but rather a gradient. Most people occupied intermediate degrees of mycophilia. Despite the remarkable similarity in the degree of mycophilia between ecological regions, the Principle-Coordinates Analysis shows differences in the specific way in which people from either region establishes a cultural relationship with mushrooms. The comparison of models suggests that sociocultural variables explains the differences better than ecological regions do. The obtained results are evidence of mycophilia among lowlands inhabitants in the Mayan region and of the fact that the mycophilia-mycophobia phenomenon is not expressed as a bimodal frequency distribution.