Differential biological activity of two extracts of ripe fruits of Carica candamarcensis Hook on human neutrophils / Actividad biologica diferencial de dos extractos de frutos maduros de Carica candamarcensis Hook en neutrófilos humanos

We evaluated the effect of the total macerate (TM) and seed oil (SO) of mature Carica candamarcensis fruits, on the release of Matrix metalloproteinase 9 (MMP9) and the phosphorylation of MAPK in neutrophils. The antioxidant capacity of these extracts was evaluated by ABTS assay. Neutrophils stimulated with different dilutions of TM or SO were analyzed for cytotoxicity, MMP9 release, and MAPK phosphorylation, using trypan blue exclusion assays, zymography, and immunoblotting, respectively. Both extracts show antioxidant activity, being higher in TM; none presented cytotoxic effect. The 5% and 2.5% dilutions of TM significantly reduced MMP9 release, and all decreased MAPK phosphorylation. SO significantly increased the release o f MMP9 and MAPK phosphorylation, the effect being greater when they were prestimulated with lipopolysaccharide.TM may have anti - inflammatory potential, while SO could have a priming effect that needs to be confirmed

Evaluamos el efecto del macerado total (MT) y aceite de semillas (AV) de frutos maduros de Carica candamarcensis , en la liberación de Matriz metaloproteinasa 9 (MMP9) y la fosfor ilación de MAPK en neutrófilos. La capacidad antioxidante de estos extractos se evaluó por ensayo ABTS. En neutrófilos estimulados con diferentes diluciones de MT o AV se analizó la citotoxicidad, liberación de MMP9 y fosforilación de MAPK, mediante ensayo s de exclusión con azul de tripano, zimografía e inmunotransferencia, respectivamente. Ambos extractos muestran actividad antioxidante, siendo mayor en MT; ninguno presentó efecto citotóxico. Las diluciones 5% y 2,5% de MT redujeron significativamente la l iberación de MMP9, y todas disminuyeron la fosforilación de MAPK. El AV incrementó significativamente la liberación de MMP9 y la fosforilación de MAPK, el efecto fue mayor cuando se preestimularon con lipopolisacárido. El MT puede tener potencial antiinfla matorio, mientras que el AV podría tener un efecto "priming" que necesita ser corroborado.

Cloning, expression and purification of fructose-2, 6-bisphosphatase gene CpF2KP in papaya / 生物工程学报

Papaya, which is mainly cultivated in the southeastern region of China, is one of the four famous fruits in Lingnan. It is favored by people because of its edible and medicinal value. Fructose-6-phosphate, 2-kinase/fructose-2, 6-bisphosphatase (F2KP) is a unique bifunctional enzyme with a kinase domain and an esterase domain that catalyzes the synthesis and degradation of fructose-2, 6-bisphosphate (Fru-2, 6-P2), an important regulator of glucose metabolism in organisms. In order to study the function of the gene CpF2KP encoding the enzyme in papaya, it is particularly important to obtain the target protein. In this study, the coding sequence (CDS) of CpF2KP, with a full-length of 2 274 bp, was got from the papaya genome. The amplified sequence of full-length CDS was cloned into the vector PGEX-4T-1 which was double digested with EcoR I and BamH I. The amplified sequence was constructed into a prokaryotic expression vector by genetic recombination. After exploring the induction conditions, the results of SDS-PAGE showed that the size of the recombinant GST-CpF2KP protein was about 110 kDa. The optimum IPTG concentration and temperature for CpF2KP induction were 0.5 mmol/L and 28 ℃, respectively. The purified sin[A1] gle target protein was obtained after purifying the induced CpF2KP protein. In addition, the expression level of this gene was detected in different tissues, and showed that the gene was expressed at the highest level in seeds and the lowest in pulp. This study provides an important basis for further revealing the function of CpF2KP protein and studying the involved biological processes of this gene in papaya.

Microclimate and development of Coffea canephora intercropped with Carica papaya: measures to mitigate climate change

Intercropped systems with Conilon coffee might provide a better environment for coffee production. The aim of this study was to assess the microclimate and development of Conilon coffee intercropped with papaya trees. Papaya was planted with spacing of 3.20 x 2.40 m. The coffee trees were planted after eight months, with spacing of 3.20 x 1.60 m, in-between papaya trees (in the same row). The measurements were taken 0, 40 and 80 cm away from the coffee plants, both in the north and south direction. Concomitantly, an adjoining full sunlight coffee system (not intercropped) was also assessed. The measurements included atmospheric parameters (temperature, irradiance, and relative humidity) and vegetative parameters for the coffee plants (leaf area, relative chlorophyll index, length of plagiotropic branches, length of orthotopic branches and number of nodes) in three periods of the year. The intercropped system of Conilon coffee and papaya trees led to a decrease in both irradiance and temperature, and higher means of relative humidity during daytime in all the periods assessed, which contributes to a better environment for coffee cultivation. The shadow provided by papaya trees in the coffee plants contributed to a higher leaf area but did not affect neither the growth of both plagiotropic and orthotopic branches, nor the number of nodes and the etiolation. The intercropped system of Conilon coffee and papaya trees may be potentially used as a farming system to mitigate climate change.

The potential use of papaya and banana peels as substrate to enhance the pigment production of Gram-positive bacterial strain isolated from Holothuria (Lessonothuria) pardalis

Aims@#The microbial pigment can be the best promising alternative to replace synthetic colorant. However, due to the high cost of synthetic medium for microbial pigment production, there is a need to develop a new low-cost medium of bacterial pigment production. This study aims to investigate the potential of banana and papaya peels as alternative lowcost substrates for a carotenoid-producing bacterium, B12 strain (bacteria strain isolated from Holothuria (Lessonothuria) pardalis).@*Methodology and results@#B12 strain identified as an aerobic bacterium with non-motile, diplobacilli shaped and Grampositive bacteria. The fermentation was optimized with different parameters included the effect of temperature, time, concentrations, pHs, carbon and nitrogen sources to find the optimum relative pigment concentration produced by B12. The results showed that the B12 strain produced the highest relative pigment concentration measured at 450 nm when the strain was cultivated at 37 °C and pH 7 in the culture medium incorporated with the combination of dried papaya peels and banana peels (100% v/v with ratio 1:1) at 72 h of incubation. Lactose, peptone and yeast were observed as the best carbon and nitrogen sources to increase the pigment concentration of B12 strain. Stability of the pigment was studied at different physiochemical stress, and it showed the pigment obtained from dried papaya and banana substrates can tolerate and stable under stress condition.@*Conclusion, significance and impact of study@#This can be concluded that the combination of dried papaya and banana peels worked well as substrate and can be utilized as a fermentation medium to replace the synthetic medium which is more expensive and uneconomical for industry application. Besides, it also helps in managing waste and solving the pollution problem due to the increasing of biochemical oxygen demand (BOD) and chemical oxygen demand (COD).

Systems biology applied to the study of papaya fruit ripening: the influence of ethylene / Biologia de sistemas aplicada ao estudo do amadurecimento de mamões: a influência do etileno

Fruit ripening is a biochemical process that results in flavor, odor, texture, and color suitable for human consumption, in addition to providing access to important nutrients. Although ripening promotes sensory and nutritional increases in fruits, there is also an increased susceptibility to physical damage, as is the case with papaya. These transformations occur due to changes in gene expression patterns at different stages of maturity, whose control and coordination result from the combined action of plant hormones, especially ethylene. As the action of this hormone in the regulation of gene expression is still elusive, this dissertation sought to address the global analysis of the transcriptome in an overview study of molecular processes involved in the ripening of ethylene-treated and non-treated papaya. Transcription factors related to ethylene synthesis and signaling had increased activity towards exogenous-ethylene treatment. Consequently, ethylene-induced enzymes had their coding genes differentially expressed, like genes related to the synthesis of carotenoids, linalool, and vitamins, which increase color, aroma, and antioxidant activity, respectively. Metabolic pathways related to the synthesis of sugars were suppressed while genes encoding the enzyme responsible for sucrose synthesis maintained a basal expression, showing that the accumulation of sugars occurs before the ripening process. The firmness of the peel and pulp of the fruits were strongly influenced by the treatment with ethylene and by the time of the experiment, suffering the action of numerous enzymes related to the degradation of the cell wall. The main enzyme responsible for softening the pulp was polygalacturonase, together with the activity of other pectinases and cellulases. In contrast to the need for the pre-climacteric action of pectate lyase and pectinesterase reported in other fleshy fruits, such as tomatoes and strawberries, papaya did not show a significant difference in their expression. The meta-analysis of several papaya ripening transcriptomes confirmed the expression profile observed in the previous RNA-seq, besides providing statistical enrichment to the biological narratives. Finally, the present study gathered a range of robust information on the gene regulation of the papaya ripening process, which opens possibilities for future approaches to transcriptomic analysis and validates the use of papaya as a model for such studies

O amadurecimento de frutos é um processo bioquímico que resulta em sabor, odor, textura e cor adequados para o consumo humano, além de propiciar o acesso a nutrientes importantes. Apesar do amadurecimento promover incrementos sensoriais e nutricionais nos frutos, ocorre também um aumento da suscetibilidade a danos físicos, como é o caso do mamão. Essas transformações ocorrem devido às alterações nos padrões de expressão gênica nos diferentes estádios de amadurecimento, cujo controle e coordenação decorrem da ação combinada de hormônios vegetais, principalmente do etileno. Como a ação deste hormônio na regulação da expressão gênica ainda é elusiva, a presente dissertação buscou abordar a análise global do transcriptoma em um amplo estudo dos processos moleculares envolvidos no amadurecimento de mamões tratados e não tratados com etileno. Os fatores de transcrição relacionados com a síntese e a sinalização do etileno tiveram sua atividade aumentada perante o tratamento exógeno com etileno. Consequentemente, as enzimas reguladas por esse hormônio tiveram seus genes de codificação expressos diferencialmente, como foi o caso de genes relacionados à síntese de carotenoides, linalool e vitaminas, que atuam no aumento da cor, aroma e atividade antioxidante, respectivamente. Vias metabólicas relacionadas com à síntese de açúcares foram reprimidas enquanto genes codificantes da enzima responsável pela síntese de sacarose mantiveram uma expressão basal, evidenciando que o acúmulo de açúcares ocorre antes do processo de amadurecimento. A firmeza da casca e da polpa dos frutos foram fortemente influenciadas pelo tratamento com etileno e pelo tempo de experimento, sofrendo ação de inúmeras enzimas relacionadas com a degradação da parede celular. A principal enzima responsável pelo amolecimento da polpa foi a poligalacturonase, em conjunto com a atividade de outras pectinases e celulases. Em contraste com a necessidade da ação pré-climatérica da pectato liase e da pectinesterase relatada em outras frutas carnosas, como tomates e morangos, o mamão não apresentou uma diferença significativa na expressão das mesmas. A meta-análise de diversos transcriptomas do amadurecimento do mamão reafirmaram o perfil de expressão observado no RNA-seq, além de prover enriquecimento estatístico às narrativas biológicas. Por fim, o presente estudo reuniu uma gama de informações robustas sobre a regulação gênica do processo de amadurecimento do mamão papaia, o que abrange a possibilidade para futuras abordagens de análise transcriptomica e valida o uso do mamão como modelo para tais estudos

Postharvest conservation of Formosa 'Tainung 1' papaya under refrigeration, modified atmosphere, and chitosan

The objective of this work is to evaluate the use of modified atmosphere and chitosan as an alternative to chemical treatment in postharvest conservation of 'Tainung 1' papaya under refrigeration. The experiment comprised of completely randomized blocks with four replications, one fruit per replication, with a 2 × 4 × 4 factorial design as follows: two types of packaging (with or without polyvinyl chloride [PVC] stretchable film, thickness of 17 µm), four treatments (control, 2% chitosan, 4% chitosan, and prochloraz fungicide with 33.75 g a.i.·100 L­1), and four storage periods (0, 7, 14, and 21 days) at 10 °C and relative humidity (RH) of 90 ± 5%. The variables evaluated were mass loss (ML), external appearance, titratable acidity (TA), soluble solids (SS), vitamin C, reducing sugars (RS), and total sugars (TS). The storage of 'Formosa' papaya under refrigeration at 10 °C and RH of 90 ± 5% associated with PVC provided a low percentage of ML and longer conservation time of apparent quality (up to 7 days) in relation to fruits with no storage in PVC. The external appearance of fruits was preserved for up to 14 days by the PVC + 4% chitosan treatment and for up to 21 days by the PVC + prochloraz fungicide treatment. The treatment PVC + 4% chitosan associated with refrigeration is efficient in maintaining fruit quality. It is, for up to 14 days, a viable alternative to the use of prochloraz fungicide for postharvest conservation of 'Formosa' papaya.

Evaluation of Caripill™ as a component of platelet storage solution

ABSTRACT Background Carica papaya Linn. has high nutraceutical and pharmacological values. The leaves possess antimicrobial, anti-tumor and antioxidant properties. They are used to treat thrombocytopenia during dengue fever and the leaf extract is commercially available as tablets under the name Caripill™ (MicroLabs, Bengaluru). Nevertheless, platelet transfusion is recommended in severe cases of thrombocytopenia, but the platelet storage is limited to 5-7 days at 22−24 °C. Reducing oxidative stress (OS) during platelet storage might help in prolonging the shelf-life, since the OS is known to cause platelet storage lesion. Hence, this study investigated the effects of Caripill™ as an additive in Tyrode's buffer during extended platelet storage. Methods Platelets isolated from 4 months old male Wistar rats were stored with Caripill™ (50 and 100 µg/ml) at 22 °C for 12 days. Platelet functional and metabolic markers and various OS markers were analyzed on days 0, 4, 8 and 12. Results Caripill™ (50 and 100 µg/ml) maintained platelet functions and lactate dehydrogenase, elevated nitrites, reduced glucose consumption, protected proteins and up-regulated the antioxidant enzymes. However, the CP100 up-regulated catalase from day 4, elevated nitrites from day 8, prevented the formation of secondary products of lipid peroxidation and increased the total antioxidant capacity on day 4. Conclusions Caripill™ reduced platelet storage lesion up to day 8 of storage. Results suggest that a higher concentration of Caripill™ was more effective in combating the oxidative damage during platelet storage. This study throws light on the beneficial effects of Caripill™ in combating oxidative stress during platelet storage.

Sensitivity of hyperparasitic fungi to alternative products for use in the control of papaya black spot / Sensibilidade de fungos hiperparasitas a produtos alternativos para uso no controle da pinta preta do mamoeiro

Abstract The use of more than one control technique can maximize the reduction of the damages caused by the fungus Asperisporium caricae causal agent of the black spot in the papaya crop. The objective of this work was to evaluate the sensitivity of the fungi Hansfordia pulvinata and Acremonium spp. to alternative products with potential for use in the control of the black-spotted ptarmigan. Three isolates of Acremonium spp. (A-598, A-602 and A-617) and an isolate of H. pulvinata (H-611) were grown in BDA medium containing Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separately. The Amistar 500WG ® fungicide was used as a positive control and pure BDA as a negative control. The toxicity of the tested products was determined based on the values of the biological index, derived from the means of mycelial growth, sporulation and germination of conidia, in each experimental unit. In this way it was possible to select the products classified as compatible for all isolates, and to test them in vivo. In the greenhouse, only the isolates and isolates with selected products, were applied in papaya plants with foliar symptoms of black-spotted. The incidence of leaves with hyperparasites and the percentage of black-painted lesions colonized by the tested isolates were evaluated. Thus, the H-611 isolate proved to be compatible with most of the alternative products tested, except with Hortifos®. Bion® and Matrix® products were compatible with all tested isolates and could be used in conjunction with Acremonium spp. and H. pulvinata to control the papaya black spot, since these products did not present toxicity on the hyperparasitic fungi.

Resumo O uso de mais de uma técnica de controle pode maximizar a redução dos danos causados pelo fungo Asperisporium caricae agente causal da pinta preta na cultura do mamoeiro. O objetivo deste trabalho foi avaliar a sensibilidade dos fungos Hansfordia pulvinata e Acremonium spp. a produtos alternativos com potencial para uso no controle da pinta-preta. Três isolados de Acremonium spp. (A-598, A-602 e A-617) e um isolado de H. pulvinata (H-611) foram cultivados em meio BDA contendo Agro-Mos®, Bion®, Ecolife®, Hortifospk®, Matriz G®, Vitaphol® separadamente. Fungicida Amistar 500WG® foi utilizado como controle positivo e BDA puro, como controle negativo. A toxicidade dos produtos testados foi determinada com base nos valores do índice biológico, provenientes das médias do crescimento micelial, esporulação e germinação de conídios, em cada unidade experimental. Dessa forma foi possível selecionar os produtos classificados como compatíveis para todos os isolados, e testá-los in vivo. Em casa de vegetação, somente os isolados e os isolados com produtos selecionados, foram aplicados em plantas de mamoeiro com sintomas foliares de pinta-preta. Avaliaram-se a incidência de folhas com hiperparasitas e a porcentagem de lesões de pinta-preta colonizadas pelos isolados testados. Assim o isolado H-611 mostrou-se compatível com a maioria dos produtos alternativos testados, exceto com Hortifos®. Os produtos Bion® e Matriz® apresentaram compatibilidade com todos os isolados testados, podendo ser utilizados em conjunto com o Acremonium spp. e H. pulvinata para controlar a pinta-preta do mamoeiro, uma vez que estes produtos não apresentaram toxidade sobre os fungos hiperparasitas.

A triple-blind, randomized controlled trial on the efficacy and safety of 1.5% Carica papaya latex cream vs. 2% Ketoconazole cream in the treatment of pityriasis versicolor among Filipinos

Background@#Carica papaya latex has been found to have antifungal properties rendering an alternative treatment for fungal infections, i.e. pityriasis versicolor. It has remarkable mycelial inhibition, and static effect on fungal growth in cultures. Its keratolytic effect can remove diseased skin cells, and enhance drug penetration. Moreover, it is organic, locally available, and relatively inexpensive.@*Objective@#To compare the efficacy and safety of 1.5% Carica papaya latex cream vs. 2% ketoconazole cream in the treatment of pityriasis versicolor among Filipinos.@*Methods@#A single-center, parallel group, triple-blind, randomized controlled trial in the Dermatology out-patient clinic of Makati Medical Center was conducted. Sixty-four patients with pityriasis versicolor were randomly allocated to the two treatment groups, and received either 1.5% Carica papaya latex cream or 2% ketoconazole cream that they used twice daily for four weeks or until cured. The participants, researcher, and assessor were blinded to the treatment assignments. Therapeutic response was assessed at weeks 1, 2, 3 and 4 based on clinical and mycologic cure. Adverse events were identified. Patients’ assessment of their improvement was done at the end of the treatment.@*Results@#All 64 subjects in both treatment groups (100% in the Carica papaya and 100% in the ketoconazole group) achieved clinical and mycologic cure within the four-week study period. The adverse reactions noted (pruritus and erythema for Carica papaya latex cream, and pruritus for ketoconazole cream) were mild, did not cause disruption of daily activities, and spontaneously resolved.@*Conclusion@#1.5% Carica papaya latex cream is an effective and safe alternative treatment to the first line therapy, ketoconazole cream, for pityriasis versicolor.

Evaluation of bioherbicidal potential of Carica papaya leaves / Avaliação do potencial de bioherbicida dase folhas de Carica papaya

Abstract Due to increased number of herbicide resistant weeds, it is needed to explore the allelopathic potential of plants as an alternative. The research was conducted to investigate allelopathic effects of Carica papaya L. leaf powder and aqueous extract on seeds as well as pre-germinated seeds of Avena fatua L., Helianthus annuus L., Rumex dentatus L., Zea mays L. and Triticum aestivum L. on filter paper and soil in Weed Management Program Laboratory, Department of Plant and Environmental Protection at PARC Institute of Advanced Studies in Agriculture, National Agriculture Research Centre, Islamabad, Pakistan. Germination percentage (%), radicle length (cm) and plumule length (cm) were parameters observed for 'Plant leaf powder bioassay' and 'Aqueous extract method'. Most significant growth inhibition was observed in A. fatua seedlings in filter paper method. A. fatua radicle length was reduced by C. papaya aqueous extract (80%) and leaf powder (89%) bioassays. Plumule length was reduced under the influence of aqueous extract (57-73%) and powdered material (59-77%). The inhibitory effects on other test species were in sequence of H. annuus followed by Z. mays and R. dentatus. The aqueous extract showed non-significant effect on wheat seed germination, radicle and plumule growth. It is suggested that C. papaya aqueous extract can be used as source of weed management in wheat crop.

Resumo Devido ao aumento do número de ervas daninhas resistentes aos herbicidas, é necessário explorar o potencial alelopático das plantas como uma alternativa. A pesquisa foi conduzida com o objetivo de investigar os efeitos alelopáticos do pó foliar de Carica papaya e do extrato aquoso das sementes, bem como das sementes pré-germinadas de Avena fatua, Helianthus annuus, Rumex dentatus, Zea mays e Triticum aestivum em papel de filtro e solo no Laboratório do Programa de Manejo de Ervas Daninhas, Departamento de Plantas e Proteção Ambiental do Instituto PARC de Estudos Avançados em Agricultura, Centro Nacional de Pesquisa Agrícola, Islamabad, Paquistão. A porcentagem de germinação (%), o comprimento radicular e o comprimento da plúmula (cm) foram os parâmetros observados para o 'Bioensaio de Pó de Folha de Planta' e o 'Método de Extração Aquoso'. A maior inibição do crescimento foi observada em mudas de A. fatua no método de papel de filtro. O comprimento radicular de A. fatua foi reduzido com os extratos aquosos de C. papaya (80%) e pó de folhas (89%). O comprimento das plúmulas foi reduzido sob a influência do extrato aquoso (57-73%) e material em pó (59-77%). Os efeitos inibitórios em outras espécies-teste foram na sequência de H. annuus seguido por Z. mays e R. dentatus. O extrato aquoso apresentou efeito não significativo na germinação das sementes de trigo, nos crescimentos radiculares e das plúmulas. Sugere-se que o extrato aquoso de C. papaya pode ser utilizado como fonte de manejo de plantas daninhas na cultura do trigo.

Oxidative foliar photo-necrosis produced by the bacteria Pseudomonas cedrina

BACKGROUND: Although bioactive metabolites capable of causing oxidative photo-necrosis in plant tissues have been identified in fungi, little is known about this type of mechanism in bacteria. These metabolites act as photosensitizers that generate reactive oxygen species (ROS) capable of causing damage to cells. In addition, these metabolites can pass into an energetically excited state when they receive some luminous stimulus, a condition in which they interact with other molecules present in the environment, such as molecular oxygen (O2), also known as triplet oxygen (3 O2), generating ROS. RESULTS: The suspension of the bacterial culture of Pseudomonas cedrina was shown to produce foliar necrosis in papaya leaves (Carica papaya L.) only in the presence of sunlight, which is evidence of photosensitizing mechanisms that generate singlet oxygen (1 O2). From the chemical study of extracts obtained from this bacteria, 3-(4-(2-carboxipropyl) phenyl) but-2-enoic acid (1) was isolated. This compound, in the presence of light and triplet oxygen (3 O2), was able to oxidize ergosterol to its peroxide, since it acted as a photosensitizer producing 1 O2, with which it was corroborated that a photosensitization reaction occurs, mechanism by which this bacterium could prove to cause oxidative foliar photo-necrosis. CONCLUSIONS: P. cedrina was able to induce oxidative foliar photo-necrosis because of its potential ability to produce photosensitizing metabolites that generate singlet oxygen in the plants it colonizes. Based on the above, it can be proposed that some bacteria can cause oxidative foliar photo-necrosis as an important mechanism in the pathogenesis of host species.

Control of anthracnose in papaya fruits by acetylsalicylic acid and 1-methylcyclopropene / Controle da antracnose em mamão por ácido acetil salicílico e 1-metilciclopropeno

This study assessed the effect of acetylsalicylic acid (ASA) and 1-methylcyclopropene (1-MCP) to control anthracnose in papaya (cultivar Golden). Disinfested-surface fruits were inoculated with Colletotrichum gloeosporioides and then the compounds were applied. The lesion diameters and the physical-chemical properties were analyzed. Assays were carried out with ASA and 1-MCP targeting the control of anthracnose and maintenance of the fruit's physical-chemical properties. The effect of ASA (20 mM; 20 min) on reducing lesion diameter occurred when applied before inoculation. Fruits treated with 1-MCP (300ppb) for 12 h showed a smaller lesion diameter than control. For the physical-chemical analysis, fruit treated with 1-MCP (200; 300 ppb; 12h) maintained fruit firmness, delayed fruit ripening and fruit fresh weight loss.

Neste trabalho avaliou-se o efeito do ácido acetilsalicílico (ASA) e 1-metilciclopropeno (1-MCP) no controle da antracnose do mamão (cv Golden). Para isso o Colletotrichum gloeosporioides foi inoculado em frutos superfícialmente desinfestados para depois aplicar-se os compostos. O diâmetro das lesões e as características físico-químicas foram determinadas. Os ensaios foram conduzidos com ASA e 1-MCP visando o controle da antracnose e a manutenção das características físico-químicas do fruto. O efeito do AAS (20 mM; 20 min) na redução do diâmetro da lesão ocorreu quando aplicado antes da inoculação. Frutos tratados com 1-MCP (300 ppb) por 12 horas apresentaram menor diâmetro de lesão que controle. Para a análise físico-química, frutos tratados com 1-MCP (200; 300 ppb; 12h) mantiveram a firmeza, o amadurecimento tardio e a perda de massa fresca dos frutos.

Antioxidant potential of Carica papaya Linn (Caricaceae) leaf extract in mice with cyclophosphamide induced oxidative stress / Potencial antioxidante do extrato de folhas de Carica papaya Linn (Caricaceae) em camundongos com estresse oxidativo induzido por ciclofosfamida

AIMS: This study aimed to investigate the effects of crude extract of Carica papaya leaves on oxidative stress in mice induced by cyclophosphamide, as well as phytochemical profile characterization of this extract. METHODS: The male Swiss mice received 15 days of treatment with the extract (500 mg kg-1, via gavage) and intraperitoneal injection of cyclophosphamide (75 mg kg-1) or saline (0.9%) on the 15th day. After 24 h the last treatment, the animals were anesthetized for blood withdrawal, sacrificed and removal of the organs for analyses (liver, kidney and heart). In the biochemical tests were determined: hematological parameters in blood, aminotransferases, alkaline phosphatase, glucose and total cholesterol dosages in plasma, enzymatic and non-enzymatic antioxidants and lipid damage marker were evaluated in different tissues, besides genotoxic and histopathological analyzes. RESULTS: In the extract of Carica papaya leaves, the flavonoids quercetin-3ß-D-glucoside and rutin were identified, besides present positive results for alkaloids, saponins and tannins. This extract increased the activity of glutathione-S-transferase and catalase enzymes in the liver and reduced the levels of reduced glutathione in the kidneys and hematocrit levels, red cell count, and hemoglobin. It promoted the decrease of the reactive species of thiobarbituric acid (TBARS) in the kidneys and the activity of enzyme aspartate aminotransferase in the plasma and was antimutagenic in the micronucleus test. CONCLUSIONS: The study showed that extract of Carica papaya was beneficial against oxidative events and prevented DNA damage. The extract also showed hepatotoxicity, therefore prolonged infusion of papaya leaves is not advisable.

OBJETIVOS: O objetivo deste estudo foi investigar os efeitos do extrato bruto de folhas de Carica papaya sobre o estresse oxidativo em camundongos induzidos pela ciclofosfamida, bem como a caracterização do perfil fitoquímico deste extrato. MÉTODOS: Os camundongos Swiss machos receberam 15 dias de tratamento com o extrato (500 mg kg-1, via gavagem) e injeção intraperitoneal de ciclofos-famida (75 mg kg-1) ou salina (0,9%) no 15º dia. Após 24 h do último tratamento, os animais foram anestesiados para retirada do sangue, sacrificados e retirada dos órgãos para análises (fígado, rim e coração). Nos testes bioquímicos foram determinados: parâmetros hematológicos em sangue, aminotransferases, fosfatase alcalina, dosagens de glicose e colesterol total no plasma, antioxidantes enzimáticos e não enzimáticos e marcador de dano lipídico foram avaliados em diferentes tecidos, além de análises genotóxicas e histopatológicas. RESULTADOS: No extrato de folhas de Carica papaya foram identificados os flavonoides quercetina-3ß-D-glicosídeo e rutina, além de resultados positivos para alcaloides, saponinas e taninos. Este extrato aumentou a atividade das enzimas glutationa-S-transferase e catalase no fígado e diminuiu os níveis de glutationa reduzida nos rins, a concentração do hematócrito, a contagem dos glóbulos vermelhos e a hemoglobina. Promoveu a diminuição das espécies reativas de ácido tiobarbitúrico (TBARS) nos rins, a atividade da enzima aspartato aminotransferase no plasma e foi antimuta-gênico no teste do micronúcleo. CONCLUSÕES: O estudo mostrou que o extrato de Carica papaya foi benéfico contra eventos oxidativos e preveniu danos no DNA. O extrato também mostrou hepatotoxicidade, portanto, a infusão prolongada de folhas de mamão não é aconselhável.

Phylogeographic distribution of asperisporium caricae isolates in brazilian southeastern and northeastern coastal regions / Distribuição filogeográfica de isolados de Asperisporium caricae no litoral do sudeste e nordeste brasileiro

Papaya (Carica papaya L.) is one of the main tropical fruits consumed in Brazil. The country is also one of the main papaya exporters, but one of the factors hindering its production lies on foliar diseases such as papaya black spot, which is caused by fungus Asperisporium caricae. This pathogen is widely distributed in the main producing regions of the Brazilian coastal area. Phylogeographic studies contribute to the knowledge about the genetic diversity and geographical distribution of genealogical lineages (haplotypes) and help better understanding the reproductive and evolutionary processes of closely related species or of a single species. Such information is useful in studies about phytopathogenic fungi because it enables identifying the most prevalent genealogical lineages in a given location, as well as inferring dispersal routes and providing information on the origin and frequency of exotic material introduction events. Results in this type of study can significantly help developing new disease control strategies. Literature still lacks studies on the Papaya x A. caricae pathosystem. Based on the phylogenetic and phylogeographic analysis applied to nucleotide sequences of the Internal transcribed spacer (ITS) gene, we herein address the genealogical and dispersal events recorded for this pathogen in order to better understand its evolution in, and adaptation to, Brazilian orchards. Three haplotypes were identified among the A. caricae isolates; their distribution was mostly related to the geographic distance between sample collection regions rather than to any reproductive or evolutionary processes presented by the species. The low variability among the herein studied isolates may result from the physiological specialization (survival exclusively associated with the host plant) and from the regional transport of contaminated fruits (with lesions and spores), besides the low contribution of reproductive events, which corroborate the lack of knowledge about the sexual stages of A. caricae.

O mamoeiro (Carica papaya L.), é uma das principais frutas tropicais consumidas no Brasil. O país é um dos principais exportadores de mamão e um dos gargalos da produção mundial está diretamente ligado à ocorrência de doenças foliares, podendo-se destacar a pinta-preta do mamoeiro, causada pelo fungo Asperisporium caricae. O patógeno está amplamente distribuído nas principais regiões produtoras, as quais englobam grande parte do litoral Brasileiro. Estudos filogeográficos contribuem não só para oconhecimento da diversidade genética e da distribuição geográfica das linhagens genealógicas (haplótipos), como também contribuem para o conhecimento dos processos reprodutivos e evolutivos de uma espécie ou de espécies estreitamente relacionadas. No estudo de fungos fitopatogênicos, tais informações são úteis para identificar linhagens genealógicas mais prevalentes em um determinado local, inferir as rotas de dispersão e fornecer informações sobre a origem e frequência dos eventos de introdução de material exótico. Este tipo de estudo produz resultados que podem colaborar significativamente na elaboração de novas estratégias de controle da doença. Não existe, até o presente momento, este tipo de estudo para o patossistema Mamoeiro x A. caricae. Neste trabalho, por meio da análise filogenética e filogeográfica, a partir de sequências nucleotídicas do gene Internal transcribed spacer (ITS), discutimos os eventos genealógicos e de dispersão deste patógeno com o intuito de compreender melhor sua evolução e adaptação nos pomares brasileiros. Dentre os isolados de A. caricae foram identificados 3 haplótipos, sendo sua distribuição relacionada mais ao distanciamento geográfico das regiões das coletas das amostras do que a eventuais processos reprodutivos ou evolutivos da espécie. Acredita-se que a baixa variabilidade dentre os isolados estudados seja explicada pela especialização fisiológica (sobrevivência exclusivamente associada à planta hospedeira) e ao transporte regional de frutos contaminados (com lesões e esporos), sendo baixa a contribuição de eventos reprodutivos, o que corrobora o desconhecimento de fase sexual de A. caricae.

Structural-dependent effects of dietary fibers in colon cancer: Focus on dietary fiber naturally changed by the papaya ripening / Efeitos estrutura-dependente das fibras alimentares no câncer de cólon: foco na fibra alimentar naturalmente modificada durante o amadurecimento do mamão papaia

ietary fiber (DF) consumption is related with several healthy benefits such as the decreasing risk of colon cancer development. The DF is not digested by the digestive enzymes and reach to colon where is fermented by the colonic microbiota. The fermentation process releases metabolites as short chain fatty acids (SCFA) such as butyrate, propionate and acetate. Besides the fermentation process, the DF can directly interact with intestinal epithelial cells inducing mechanism that can also be related with the associated DF consumption benefits. The lack of information regarding DF and colon cancer are due to the complexity of both the cancer and the DF structure. The papayas DF are derived from the fruit cell wall, and they are probably naturally modified during ripening through a massive polysaccharide hydrolysis, because papayas show a very fast pulp softening. Due to the lack of information about DF and their beneficial effects to human health as well as the possibility of the natural papaya ripening to modifying the DF presented in the fruit pulp, the present thesis had as the primary objectives: 1) to evaluate how the cell-wall degrading enzymes affect the fruit cell wall solubilization and molecular weight; 2) to investigate the direct effects of the papaya pectin derived from unripe to ripe papayas in cancer cell lines, in galectin-3 interaction and in HEK cells expressing pattern recognition receptors (PRR); 3) to evaluate the human colonic in vitro fermentation using DF from unripe and ripe papayas as substrates; 4) to conduct an in vivo experiment using rats with pre-neoplastic colon lesions while receiving a diet with DF from unripe and ripe papayas. The endopolygalacturonases were the main enzymes acting on the solubilizing papaya cell wall pectin affecting both the papaya firmness and pectin structure. Overall, the papayas DF showed a ripening dependent structureeffects. In the cancer cell lines experiments, the ripe papayas pectin showed a more pronounced effects in inducing cancer cell death, inhibiting cancer cells migration and aggregation, activating PRR as toll-like receptors and inhibiting the pro-metastatic protein galectin-3. The DF from papayas also showed different aspects in colonic in vitro fermentation regarding the DF utilization by the bacteria and the bacteria abundance profile. Lastly, the animals receiving the diet with the DF from ripe papayas had less aberrant crypt foci in colon than the animals that received the DF from unripe papayas or cellulose (AIN-93G DF). Therefore, the study of papaya DF was carried out both during papaya ripening and its biological effects in vitro and in vivo, generating unprecedented results relating the endogenous biochemical changes of the fruits during maturation with the possible beneficial effects of their ingestion for health human

O consumo de fibras alimentares (FA) está relacionado com vários benefícios à saúde como a diminuição no risco do desenvolvimento de câncer de cólon. A FA não é digerida pelas enzimas digestivas do trato gastrointestinal sendo fermentada pela microbiota intestinal do cólon. Como subproduto do processo de fermentação há a liberação de ácidos graxos de cadeia curta (SCFA) - como o butirato, o propionato e o acetato. Além do processo de fermentação, a FA pode interagir diretamente com as células epiteliais do intestino, induzindo mecanismos que também podem estar relacionados com os benefícios associados ao consumo de FA. A falta de informação sobre a FA e o câncer de cólon é, em partes, devido à complexidade de ambos, tanto do câncer quanto da estrutura da FA. As FA do mamão papaia são derivadas da parede celular da fruta apresentando diferentes estruturas dependendo do ponto de amadurecimento do fruto. Esse fato ocorre, pois, durante o amadurecimento do mamão papaia, existe uma extensa hidrólise dos polissacarídeos presentes na parede celular, diminuindo rapidamente a firmeza da polpa do fruto. Devido à falta de informações sobre FA e seus efeitos benéficos à saúde humana que são dependentes da sua estrutura, bem como a possibilidade do amadurecimento do mamão papaia naturalmente modificar as FA presentes na polpa dos frutos, a presente tese teve como principais objetivos: 1) avaliar como as enzimas que degradam a parede celular do mamão papaia afetam a solubilização e o peso molecular da parede celular do fruto; 2) investigar os efeitos diretos da pectina derivada de mamões verdes e maduros em linhagens de células de câncer, na interação com a galectina-3, e em células do tipo HEK que expressam receptores de reconhecimento de padrões (RRP); 3) avaliar a fermentação colônica humana in vitro utilizando as FA de mamões verdes e maduros; 4) avaliar em ratos com lesões pré-neoplásicas no cólon o efeito do consumo de ração com ou sem FA de mamões papaias verdes e maduros. As endopoligalacturonases foram relacionadas como as principais enzimas que atuam solubilizando a pectina da parede celular do mamão, afetando tanto a firmeza da polpa do fruto quanto a solubilização da pectina durante o amadurecimento. De modo geral, as FA dos mamões exerceram um efeito estruturadependente de acordo com a maturação do fruto. Nos experimentos utilizando linhagens de células de câncer, a pectina do mamão papaia maduro apresentou efeitos mais pronunciados na indução da morte e na inibição da migração e da agregação das células, bem como ativando os RRP, como por exemplo, os receptores do tipo toll-like, além de inibir a proteína pró-metastática galectina-3. As FA dos mamões também apresentaram diferentes resultados na fermentação colônica in vitro quanto à utilização das FA pelas bactérias do intestino, e também no perfil de crescimento dessas bactérias. Por fim, os animais que receberam a dieta com as FA dos mamões maduros apresentaram menor incidência de focos de criptas aberrantes do que os animais que receberam as FA provenientes de mamões verdes ou de celulose (FA da ração AIN-93G). Portanto, o estudo das FA dos mamões foi efetuado tanto durante o amadurecimento dos mamões quanto dos seus efeitos biológicos in vitro e in vivo, tendo gerado resultados inéditos relacionando as alterações bioquímicas endógenas dos frutos durante o amadurecimento com os possíveis efeitos benéficos da sua ingestão para a saúde humana

Oviposition of fruit flies (Diptera: Tephritidae) and its relation with the pericarp of citrus fruits / Oviposição de moscas-das-frutas (Diptera: Tephritidae) e sua relação com o pericarpo de frutos cítricos

Abstract Fruit flies (Diptera: Tephritidae) represent a threat to fruit growing worldwide, mainly the citrus culture, however, biological studies show that fruit flies are not perfectly adapted to this host. This study investigated oviposition of Anastrepha fraterculus (Wiedemann, 1830) and Ceratitis capitata (Wiedemann, 1824) and its relation with the pericarp of citrus fruits. We evaluated the relationship between depth of oviposition of A. fraterculus and C. capitata and epicarp thickness of orange [Citrus sinensis (L.) Osbeck)] 'Navelina' and tangerine [C. reticulata (L.)] 'Clemenules' and the influence of fruit mesocarp of tangerine 'Clemenules' on oviposition of these species. The study was conducted under controlled conditions of temperature (25 ± 2 °C), relative humidity (70 ± 10% RH) and photophase (12 h). A. fraterculus and C. capitata laid their eggs in the flavedo region of orange 'Navelina' and between the albedo and flavedo of tangerine 'Clemenules'. When fruits with mesocarp exposed were offered, there was no oviposition by both fruit fly species. The results show that epicarp thickness of citrus fruits did not influence oviposition of A. fraterculus and C. capitata as oviposition did not occur only in the presence of the mesocarp, suggesting that other factors are involved in oviposition of these species.

Resumo As moscas-das-frutas (Diptera: Tephritidae) representam um risco à fruticultura mundial, especialmente na cultura dos citros, entretanto estudos biológicos demonstram que as moscas-das-frutas não estão perfeitamente adaptadas à estes hospedeiros. Este estudo investigou a oviposição de Anastrepha fraterculus (Wiedemann, 1830) e Ceratitis capitata (Wiedemann, 1824) e sua relação com o pericarpo de frutos cítricos. Foi avaliada a relação entre a profundidade de oviposição de A. fraterculus e de C. capitata e a espessura do epicarpo dos frutos de laranjeira [Citrus sinensis (L.) Osbeck)] 'Navelina' e tangerineira [C. reticulata (L.)] 'Clemenules' e a influência do mesocarpo de frutos de tangerineira 'Clemenules' na oviposição destas espécies. O estudo foi conduzido em condições controladas de temperatura (25 ± 2 °C), umidade relativa (70 ± 10%) e fotofase (12 horas). A. fraterculus e C. capitata depositaram ovos no flavedo de frutos de laranjeira 'Navelina' e entre o flavedo e o albedo de frutos de tangerineira 'Clemenules'. Quando oferecido frutos com mesocarpo exposto, não houve oviposição por ambas as espécies de mosca. Os resultados demonstram que a espessura do epicarpo de frutos cítricos não influenciou a oviposição de A. fraterculus e de C. capitata, a qual não ocorreu na presença apenas do mesocarpo, sugerindo que outros fatores estão envolvidos na oviposição por estas espécies.

Control of Phytophthora palmivora on postharvest papaya with Trichoderma asperellum, T. virens, T. harzianum and T. longibrachiatum / Controle de Phytophthora palmivora em mamão na pós-colheita por Trichoderma asperellum, T. virens, T. harzianum e T. longibrachiatum

Papaya (Carica papaya) is one of the most cultivated and consumed tropical fruit worldwide. Its production might be limited by preharvest and postharvest diseases. The fruit rot caused by Phytophthora palmivora is one of the most important postharvest diseases of papaya in Brazil. The control of these diseases is usually made with fungicide applications. Therefore, studies concerning biocontrol of postharvest diseases might generate data that may reduce the environmental impacts caused by pesticides. Thus, the biological control by Trichoderma in postharvest diseases is an alternative to the use of fungicides for the postharvest control of P. palmivora in the papaya fruit. Four antagonists [T. asperellum (SF04), T. virens (255C1), T. harzianum (THP) and T. longibrachiatum (4088)] were tested, as follow: 1) Trichoderma spp. applied 1 hour after inoculation of P. palmivora and; 2) Trichoderma spp. applied 24 hours after inoculation of P. palmivora; 3) Trichoderma spp. applied 1 hour before inoculation of P. palmivora, and 4) Trichoderma spp. applied 24 hours before inoculation of P. palmivora. All Trichoderma significantly (P£0,05) reduced the incidence and severity of disease. The 4088 (T. longibrachiatum) isolate was the best controller agent of P. palmivora in postharvest.

O mamão é uma fruta muito cultivada e consumida nas regiões tropicais e subtropicais do mundo e apresenta diversos problemas fitossanitários. Sendo assim, estudos de doenças pós-colheita com biocontroladores viabilizam a diminuição de impactos causados pelo uso de fungicidas. A podridão-dos-frutos (Phytophthora palmivora) é uma importante doença pós-colheita em mamão no Brasil. Neste contexto, o controle biológico desta doença na póscolheita com Trichoderma é uma alternativa viável ao uso de fungicidas e foi aplicado neste estudo para avaliar a eficácia de Trichoderma spp. para o biocontrole de P. palmivora em mamão na pós-colheita. Foram utilizados quatro potenciais antagonistas: T. asperellum (SF04), T. virens (255C1), T. harzianum (THP) e T. longibrachiatum (4088). E as frutas foram submetidas aos seguintes tratamentos: Inoculação de P. palmivora e 1 hora depois inoculação do Trichoderma spp.; Inoculação de P. palmivora e 24 horas depois inoculação do Trichoderma spp.; Inoculação de Trichoderma spp. e 1 hora depois inoculação do P. palmivora e; Inoculação de Trichoderma spp. e 24 horas depois inoculação do P. palmivora. Todos os isolados de Trichoderma reduziram significativamente tanto na incidência como na severidade da doença. O isolado 4088 (T. longibrachiatum) foi o melhor no controle da podridão.

Reduction of papaya rot (Phytophthora palmivora) with phosphite and Acibenzolar-S-Methyl in preharvest and postharvest / Redução da podridão do mamão (Phytophthora palmivora) com fosfitos e Acibenzolar-S-Metil em pré e pós-colheita

The papaya fruit rot (Phytophthora palmivora) is responsible for significant losses. To reduce diseases, especially in areas with climate and humidity favorable to pathogens, are adopted chemical methods, which sometimes increase the cost of production and cause severe environmental impacts. Alternatively, there are products, such as, phosphites of potassium and acibenzolar-S-methyl (ASM) that might be efficient on disease control and less aggressive to environment. Phosphites of K and ASM were evaluated in this study on the control effectiveness of papaya fruit rot at different dosages in preharvest and postharvest. The severity and percentage of disease control were evaluated for each treatment. For the pre-harvest treatments (applied six days before harvest), the phosphite of K [240 g L-1 K2O, 340 g L-1 P2O5 and 50 g L-1 (Reforce® + Salicylic Acid)] at 3 or 6 mL L-1significantly reduced disease severity, and, reduced fruit ripening. On postharvest application, ASM reduced disease severity.

A podridão dos frutos do mamoeiro (Phytophthora palmivora) é responsável por perdas significativas e para minimizar a doença, principalmente em locais com clima favorável ao patógeno, medidas de controle químico são adotadas. Porém, estas medidas podem elevar o custo da produção e causar severos impactos ambientais. Alternativamente, existem produtos como os fosfitos de potássio (K) e acibenzolar-S-metil (ASM) que podem ser eficientes no controle de doenças e menos agressivos ao ambiente. Fosfitos de K e ASM foram avaliados neste trabalho quanto à eficácia do controle da podridão de frutos de mamão 'Sunrise Solo' tipo exportação, cultivados no Sul da Bahia, em diferentes doses e formulações comerciais na pré e pós-colheita. Para cada tratamento foi avaliada a severidade da doença e o percentual de controle da doença, sendo também verificada a influência nas características fisioquímicas na fruta. Para os tratamentos pré-colheita, aplicados aos seis dias antes da colheita, o fosfito de K [240 g L-1 K2O, 340 g L-1 P2O5 e 50 g L-1 C7H6O3 (Reforce® + Ácido Salicílico)] a 3 ou 6 mL L-1 reduziu significativamente a doença, e influenciou diretamente no atraso da maturação da fruta. Na pós-colheita, ASM incitou redução na severidade da doença. Todos os tratamentos não alterarão as características fisioquímicas da fruta.

Evaluation of papaya genotypes using agronomic descriptors and estimation of genetic parameters / Descritores agronômicos e parâmetros genéticos na avaliação de genótipos de mamoeiros

In Brazil, the improvement of papaya agribusiness is correlated to the estimation of genetic parameters and evaluation of agronomic descriptors; thus enabling less favourable areas for the development and implementation of papaya crops. Therefore, the current study aimed to evaluate quantitative descriptors and to estimate genetic parameters in genotypes of papaya grown under subtropical climate in the state of São Paulo. The experimental design was randomized in block. The experimental units were grouped into seven blocks of five plants each and three treatments, which consisted of three different varieties: Sunrise Solo, Tainung nº1 and local. The psychochemical properties were analysed for plant height; intersection height of first fruits; stem diameter; male, female and hermaphroditic flower percentage; fruit number per plant; production; yield; normal fruits percentage, carpeloid and pentandric fruits; pulp matter; pulp length; pulp diameter; pulp shape; pulp cavity; pulp thickness; peel performance; pulp performance; seeds performance; seeds number; soluble solids; pH; titratable acidity; ratio; technological index; ascorbic acid and reducing sugars. Furthermore, most agronomic descriptors showed high heritability; besides that, the varieties Sunrise Solo and Tainung nº1 performed the best results in Botucatu.

A estimativa de parâmetros genéticos e a avaliação de descritores agronômicos do mamoeiro pode contribuir com o desenvolvimento do agronegócio do mamão no Brasil, permitindo a expansão de novos cultivos em regiões menos favoráveis para sua exploração comercial. Frente ao exposto, objetivou-se com este trabalho avaliar descritores quantitativos e estimar parâmetros genéticos em genótipos de mamoeiro, cultivados em clima subtropical do estado de São Paulo. O delineamento experimental foi em blocos casualizados, com três tratamentos (variedade Sunrise Solo, híbrido Tainung nº1 e variedade local) e sete blocos, utilizando cinco plantas por parcela experimental. Foram mensuradas as seguintes caraterísticas: altura das plantas e da interseção dos primeiros frutos; diâmetro do caule; porcentagens de plantas masculinas, femininas e hermafroditas; número de frutos por planta; produção; produtividade; porcentagem de frutos normais, carpelóides e pentândricos; massa, comprimento, diâmetro, formato, diâmetro da cavidade interna e espessura da polpa dos frutos; rendimento de casca, de polpa e de sementes; número de sementes; sólidos solúveis; pH; acidez titulável; ratio; índice tecnológico; ácido ascórbico e açúcares redutores. Foi possível inferir com o estudo que há alta herdabilidade quanto à maioria dos descritores agronômicos avaliados e que os bons resultados para a variedade Sunrise Solo e o híbrido Tainung nº 1, possibilitam a indicação desses genótipos para cultivo, na região de Botucatu ­ SP.