Biotechnology applied to Annona species: a review

Annonaceae is an ancient family of plants including approximately 50 genera growing worldwide in a quite restricted area with specific agroclimatic requirements. Only few species of this family has been cultivated and exploited commercially and most of them belonging to the genus Annona such as A. muricata, A. squamosa, the hybrid A. cherimola x A. squamosa and specially Annona cherimola: the cherimoya, commercially cultivated in Spain, Chile, California, Florida, México, Australia, Ecuador, Peru, Brazil, New Zealand and several countries in South and Central America. The cherimoya shows a high degree of heterozygosis, and to obtain homogeneous and productive orchards it is necessary to avoid the propagation by seeds of this species. Additionally, the traditional methods of vegetative propagation were inefficient and inadequate, due to the low morphogenetic potential of this species, and the low rooting rate. The in vitro tissue culture methods of micropropagation can be applied successfully to cherimoya and other Annona sp to overcome these problems. Most of the protocols of micropropagation and regeneration were developed using the cultivar Fino de Jete, which is the major cultivar in Spain. First it is developed the method to micropropagate the juvenile material of cherimoya (ENCINA et al., 1994), and later it was optimized a protocol to micropropagate adult cherimoya genotypes selected by outstanding agronomical traits (PADILLA and ENCINA, 2004) and further it was improved the process through micrografting (PADILLA and ENCINA, 2011).At the present time we are involved in inducing and obtaining new elite genotypes, as part of a breeding program for the cherimoya and other Annonas, using and optimizing different methodologies in vitro: a) Adventitious organogenesis and regeneration from cellular cultures (ENCINA, 2004), b) Ploidy manipulation of the cherimoya, to obtain haploid, tetraploid and triploid plants (seedless), c) Genetic transformation, for the genes introduction to control the postharvest processes and the genes introduction to provide resistance to pathogen and insects and d) Micropropagation and regeneration of other wild Annona or related Annonaceae species such as: Annona senegalensis, A. scleroderma, A. montana, A. reticulata, A. glabra, A. diversifolia and Rollinia sp.

Underutilized Annona Species from the Brazilian Cerrado and Amazon Rainforest: A Study on Fatty Acids Profile and Yield of Seed Oils

Annona (Annonaceae) is an important source of fruits in the Brazilian Cerrado and the Amazon Rainforest. Some Annona species are widely commercialized as fresh fruit or as frozen pulp. Seeds are accustomedly discarded. Our main goal was to analyze fatty acids profile from seeds of A. crassiflora and A. coriacea from Cerrado, A. montana from Amazon Forest, and three cultivars (A. cherimola cv. Madeira, A. cherimola x A. squamosa cv. Pink`s Mammonth and A. cherimola x A. squamosa cv. Gefner). The total oil yield ranged between 20 and 42% by weight of dry mass. The A cherimola x A. squamosa cv. Gefner has significantly higher total lipid yield than all other samples. 100 g of fruit of this species present 6-8 g of seeds. Considering the fruit production of Chile (over 221 ton of fruits/year), more than 1300 ton of seed/year could be obtained, which could provide at least 200 ton of seed oil. Oleic acid was predominant for most samples, but for A. montana linoleic acid was the most abundant FA. Phenotypic variation on FAME profile was observed. These new data are an urgent requirement for supporting conservation programs, mainly for Cerrado areas in Brazil.

Determinação da qualidade e do teor de sólidos solúveis nas diferentes partes do fruto da pinheira (Annona squamosa L.)

O objetivo foi avaliar algumas características qualitativas e estimar o teor de sólidos solúveis (TSS) nas porções basal (ligada ao pedúnculo), mediana e apical do fruto da pinheira. Para a estimação desse teor, três pinhas de cada uma de dez árvores, de pomares localizados em Mossoró-RN e Aracati-CE, com idade entre seis e oito anos, foram utilizadas. As árvores foram consideradas repetições (blocos), os frutos foram considerados parcelas e as porções dos frutos, subparcelas. Três frutos de cada árvore foram descascados e divididos em três porções aproximadamente iguais, da base para o ápice. O material foi triturado e filtrado. No suco obtido, foram feitas três leituras em um refratômetro. A média destas três leituras representou o valor de cada subparcela. A análise de variância conjunta indicou efeitos de locais, blocos em locais, porções e da interação locais x frutos x porções. O peso do fruto variou de 226 a 418 g e o rendimento de polpa, de 45 a 54 %. O TSS na porção basal (26,49 %) foi inferior aos teores das porções mediana (28,02 %) e apical (27,53 %), mas não diferiu significativamente daquele da porção apical.

Uso de cera e 1-metilciclopropeno na conservação refrigerada de graviola (Annona muricata L.)

Graviolas 'Morada', provenientes de pomar comercial localizado em Limoeiro do Norte, Ceará, foram colhidas na maturidade fisiológica com o objetivo de avaliar o efeito da aplicação pós-colheita de 1-meticiclopropeno (1-MCP) e cera na conservação, durante o armazenamento refrigerado. Os frutos foram armazenados por 0; 4; 8; 11; 13 e 15 dias, a 15,4±1,1ºC e 86,0±7,3% UR. Os tratamentos pós-colheita foram os seguintes: controle, 200 nL L-1 de 1-MCP (SmartFresh™), pulverização com a cera Fruit wax® e pulverização com Fruit wax® seguida de aplicação de 200 nL L-1 de 1-MCP. O delineamento experimental foi o inteiramente casualizado, em fatorial 4x6 (tratamentos pós-colheita x tempo de armazenamento), com quatro repetições. As variáveis analisadas foram: cor da casca e da polpa, a partir da luminosidade (L), cromaticidade (C) e ângulo de cor (Hº); perda de massa; pH; acidez titulável (AT); sólidos solúveis totais (SST); açúcares solúveis totais (AST) e açúcares redutores (AR). Os tratamentos cera e cera+1-MCP mantiveram estáveis os valores de L da casca e da polpa até o 8º e o 4º dia de armazenamento, respectivamente. O aumento em SST foi temporariamente atrasado pelos tratamentos pós-colheita. A AT, pH, AST e AR não foram influenciados pela cera e pelo 1-MCP. O uso da cera diminuiu a perda de massa em 23%, quando comparado ao controle.

Influência da temperatura na germinação de sementes de Annona Montana

Com o objetivo de avaliar a influência da temperatura sobre a porcentagem de germinação e o índice de velocidade de germinação (IVG) de sementes de Annona montana, conhecida como falsa graviola, espécie com potencial para porta-enxerto das variedades comerciais, testaram-se quatro temperaturas (20; 25; 30 e 35ºC). O trabalho foi desenvolvido em laboratório em câmaras de germinação tipo BOD, utilizando-se de sementes de três plantas provenientes do Banco de Germoplasma do Departamento de Produção Vegetal da Faculdade de Ciências Agrárias e Veterinárias - UNESP, Câmpus de Jaboticabal-SP. O delineamento experimental foi o inteiramente casualizado, com quatro repetições, com dez sementes cada. Pelos resultados obtidos, tem-se que tanto para o parâmetro porcentagem de germinação como para o IVG, os maiores valores observados foram para as sementes na temperatura de 30ºC (55% de germinação e IVG = 0,153), seguido da temperatura de 25ºC (25% de germinação e IVG = 0,088). Para as temperaturas de 20ºC e 35ºC, não foi observada ocorrência de germinação. A análise estatística dos dados de porcentagem de germinação foi transformada em arc-sen square root (x/100) e as médias foram comparadas pelo teste de Tukey, a 5% de probabilidade.

Influência da temperatura na germinação de sementes de Annona Montana

Com o objetivo de avaliar a influência da temperatura sobre a porcentagem de germinação e o índice de velocidade de germinação (IVG) de sementes de Annona montana, conhecida como falsa graviola, espécie com potencial para porta-enxerto das variedades comerciais, testaram-se quatro temperaturas (20; 25; 30 e 35ºC). O trabalho foi desenvolvido em laboratório em câmaras de germinação tipo BOD, utilizando-se de sementes de três plantas provenientes do Banco de Germoplasma do Departamento de Produção Vegetal da Faculdade de Ciências Agrárias e Veterinárias - UNESP, Câmpus de Jaboticabal-SP. O delineamento experimental foi o inteiramente casualizado, com quatro repetições, com dez sementes cada. Pelos resultados obtidos, tem-se que tanto para o parâmetro porcentagem de germinação como para o IVG, os maiores valores observados foram para as sementes na temperatura de 30ºC (55% de germinação e IVG = 0,153), seguido da temperatura de 25ºC (25% de germinação e IVG = 0,088). Para as temperaturas de 20ºC e 35ºC, não foi observada ocorrência de germinação. A análise estatística dos dados de porcentagem de germinação foi transformada em arc-sen square root (x/100) e as médias foram comparadas pelo teste de Tukey, a 5% de probabilidade.

Annona sp: Plants with multiple applications as alternative medicine - a review

Annona species have been used as a natural remedy for a variety of illnesses with antiparasitic, antispasmodic, antidiarrheal, antiulcer, sedative, analgesic, hypotensive, and vermifugal effects. These properties are due to the presence of a number of bioactive compounds on the leaves, fruit, seeds, and stem. The aim of this review is to show the main species of Annona, their medicinal properties and the chemical constituents that may be related to these effects. In the leaves it is possible to find acetogenins, annopentocins A, B, and C, cis- and trans-annomuricin-D-ones, goniothalamicin, arianacin, and javoricin, related to anticancer properties. Quercetin-3-O-glucoside, also found in the leaves mediates antidiabetic and antioxidative effects. In the fruit are found annonaine, nornuciferine and asimilobine, associated to antidepressive effects. In the seeds are found muricatetrocin A and muricatetrocin B, longifolicin, corossolin, corossolone, uvarigrandin A, bullatacin, squamotatin. These acetogenins are associated with anticancer effects. Cyclosquamosin B, quercetin, and cyclosquamosin from the seeds have respectively vasorelaxant, antithyroidal and, antiinflammatory activity. In the stem parts there are several components as N-trans-feruloyltyramine, N-p-coumaroyltyramine, and N-trans-caffeoyltyramine, lignans, syringaresinol, syringaldehyde, beta-sitosterol and beta-sitosterol-beta-D-glucoside which exhibit antiplatelet aggregation activity. Copaene, patchoulane, 1H-cycloprop (e) azulene and kaur-16-en-19-oic acid found in the barks exhibit significant central as well as peripheral analgesic and antiinflammatory activities. The properties of the biological compounds in Annona species support information that may provide validation for its medicinal uses, but further studies should be performed to establish ideal and safe doses of consumption to ensure the effectiveness of the benefits. © 2012 Bentham Science Publishers.

Condicionamento osmótico e GA3 na germinação de sementes de araticum de terra fria (Annona emarginata (Schltdl.) H. Rainer)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Reguladores vegetais na superação da dormência, balanço hormonal e degradação de reservas em sementes de Annona diversifolia SAFF. e A. purpurea Moc. & Sessé Ex Dunal (Annonaceae)

As sementes de anonáceas são conhecidas por apresentarem mecanismos de dormência, o que dificulta a perpetuação das espécies e a formação de áreas produtivas para a exploração comercial. Deste modo, os objetivos deste trabalho foram estudar curva de aquisição de água; a germinação de sementes tratadas com GA3 e GA4+7 + Benziladenina; o balanço hormonal e a degradação de reservas em sementes de Annona diversifolia Saff e Annona purpurea Moc & Sessé ex Dunal tratadas com reguladores vegetais para a superação da dormência. Para tanto foram realizados três experimentos. Para a construção da curva de aquisição de água foram utilizadas 4 repetições de 25 sementes que foram mantidas em embebição e pesadas durante 480 horas. O segundo experimento foi constituído pela germinação das sementes tratadas com os reguladores vegetais; o delineamento experimental empregado foi o inteiramente casualizado com 4 repetições de 25 sementes por parcela em esquema fatorial 2 x 7 (reguladores x concentrações). Os tratamentos foram constituídos pelas combinações entre concentrações de GA3 e de GA4+7 + Benziladenina (GA4+7 + BA) x 0, 200, 400, 500, 600, 800 e 1000 mg L-1 i.a.. No terceiro experimento foram quantificados ABA (Ácido abscísico) e GA (Giberelinas), proteínas, açúcares...

Florivory and sex ratio in Annona dioica St. Hil. (Annonaceae) in the Pantanal at Nhecolândia, southwestern Brazil

Annona dioica St. Hil. is a species that grows to approximately 2 m tall and is very widespread in the cerrados. Individual plants of this androdioecious species produce numerous hermaphroditic or male flowers, but few fruits. The aim of this study was to determine the sex ratio among the plants and to compare the frequency of herbivory between male and hermaphroditic flowers. The fieldwork was done by studying flowering plants in grasslands used as pasture for cattle at Fazenda Nhumirim. One hundred and forty-seven male plants and 71 hermaphroditic plants were examined and produced a total of 194 and 94 flowers, respectively, during the study period. The male:hermaphrodite sex ratio was 2.07:1, and was similar to the male:hermaphrodite flower ratio of 2.06:1. The frequency of florivory rate in hermaphrodites was significantly higher than in male flowers (33.0%, n = 31, and 25.7%, n = 50, respectively; G = 14.83; d.f. = 1; p < 0.001). The mean fresh weights of male and hermaphroditic flowers were significantly different (8.38 ± 2.40 g vs. 6.93 ± 2.68 g, respectively; 0 ± SEM; n = 50 each; t = 2.479; d.f. = 49; p = 0.017). These results indicate that the low fruit set in this species can be explained by the sex ratio, the greater herbivory of hermaphroditic flowers and the probable absence of pollinators.

Chemical constituents of the volatile oil from leaves of Annona coriacea and in vitro antiprotozoal activity

The essential oil of the leaves from Annona coriacea Mart., Annonaceae, was extracted by hydrodistillation in a Clevenger apparatus and analyzed by GC/MS and GC/FID. The oil yield was 0.05% m/m. Sixty compounds were identified, in a complex mixture of sesquiterpenes (76.7%), monoterpenes (20.0%) and other constituents (3.3%). Bicyclogermacrene was its major compound (39.8%) followed by other sesquiterpenes. Most of the monoterpenes were in low concentration (<1%). Only β-pinene and pseudolimonene presented the highest level of 1.6%. The volatile oil presented anti-leishmanial and trypanocidal activity against promastigotes of four species of Leishmania and trypomastigotes of Trypanosoma cruzi, showing to be more active against Leishmania (L.) chagasi (IC50 39.93 µ g/mL) (95% CI 28.00-56.95 µ g/mL).

Relationship between fruit traits of custard apple trees (Annona squamosa L.)

The objective of this study was to estimate simple and partial coefficients of correlation, as well as to divide their effects into direct and indirect using path analysis for custard apple tree traits. Twenty half-sibling progenies were evaluated in a randomized block design with five replicates, and plots consisting of four plants. Six traits were evaluated in the first cropping season (mean number of seeds per fruit and mean weight of the pericarp, pulp, pedicel, seeds per fruit, and the whole fruit), while five traits were evaluated in the first three cropping seasons (mean fruit length and width, total number of fruits ha-1, mean fruit weight (in both types of analyses), and fruit yield in kg ha-1). The results of this work led to the conclusion that doing selection based on simple correlation estimates may not be convenient, since not always a cause and effect relationship can be verified between two traits. Positive correlations were obtained between number of seeds and seed weight, and between number of fruits and yield. The greatest direct effects were those obtained for pulp weight on fruit weight and for mean number and weight of fruits on fruit yield. The most important indirect effects were obtained for number of seeds and pericarp weight, obtained via pulp weight, on fruit weight, and for fruit length and width, obtained via mean fruit weight, on fruit yield.

Genetic diversity in sugar apple (Annona squamosa L.) by using RAPD markers

Genetic diversity in a collection of 64 sugar apple accessions collected from different municipalities in northern Minas Gerais was assessed by RAPD analysis. Using 20 selected RAPD primers 167 fragments were generated, of which 48 were polymorphic (28.7%) producing an average of 2.4 polymorphic fragments per primer. Low percentage of polymorphism (< 29%) was observed by using the set of primers indicating low level of genetic variation among the 64 accessions evaluated. Genetic relationships were estimated using Jaccard's coefficient of similarity. Accessions from different municipalities clustered together indicating no correlation between molecular grouping and geographical origin. The dendrogram revealed five clusters. The first cluster grouped C19 and G29 accessions collected from the municipalities of Verdelândia and Monte Azul, respectively. The second cluster grouped G16 and B11 accessions collected from the municipalities of Monte Azul and Coração de Jesus, respectively. The remaining accessions were grouped in three clusters, with 8, 15 and 37 accessions, respectively. In summary, RAPD showed a low percentage of polymorphism in the germplasm collection.

Antibacterial effect (in vitro) of Moringa oleifera and Annona muricata against Gram positive and Gram negative bacteria

Antibacterial effects of aqueous and ethanolic extracts of seeds of moringa (Moringa oleifera) and pods of soursop (Annona muricata) in the concentration of 1:5 and 1:10 in volumes 50, 100, 150 and 200 µL were examined against Staphylococcus aureus, Vibrio cholerae, Escherichia coli (isolated from the organism and the aquatic environment) and Salmonella Enteritidis. Antibacterial activity (inhibition halo > 13 mm) against S. aureus, V. cholerae and E. coli isolated from the whiteleg shrimp, Litopenaeus vannmaei, was detected in aqueous and ethanolic extracts of moringa. E. coli isolated from tilapiafish, Oreochromis niloticus, was sensitive to the ethanolic extract of moringa. The aqueous extracts of soursop showed an antibacterial effect against S. aureus and V. cholerae, but the antibacterial activity by the ethanol extracts of this plant was not demonstrated.

In vitro SCREENING ANTIBACTERIAL ACTIVITY OF Bidens pilosa LINNÉ AND Annona crassiflora MART. AGAINST OXACILLIN RESISTANT Staphylococcus aureus (ORSA) FROM THE AERIAL ENVIRONMENT AT THE DENTAL CLINIC

Currently multiresistant Staphylococcus aureus is one common cause of infections with high rates of morbidity and mortality worldwide, which directs scientific endeavors in search for novel antimicrobials. In this study, nine extracts from Bidens pilosa (root, stem, flower and leaves) and Annona crassiflora (rind fruit, stem, leaves, seed and pulp) were obtained with ethanol: water (7:3, v/v) and their in vitro antibacterial activity evaluated through both the agar diffusion and broth microdilution methods against 60 Oxacillin Resistant S. aureus (ORSA) strains and against S. aureus ATCC6538. The extracts from B. pilosa and A. crassiflora inhibited the growth of the ORSA isolates in both methods. Leaves of B. pilosa presented mean of the inhibition zone diameters significantly higher than chlorexidine 0.12% against ORSA, and the extracts were more active against S. aureus ATCC (p < 0.05). Parallel, toxicity testing by using MTT method and phytochemical screening were assessed, and three extracts (B. pilosa, root and leaf, and A. crassiflora, seed) did not evidence toxicity. On the other hand, the cytotoxic concentrations (CC50 and CC90) for other extracts ranged from 2.06 to 10.77 mg/mL. The presence of variable alkaloids, flavonoids, tannins and saponins was observed, even though there was a total absence of anthraquinones. Thus, the extracts from the leaves of B. pilosa revealed good anti-ORSA activity and did not exhibit toxicity.