Mutagénesis inducida en plantas de Solanum betaceum Cav. mediante el uso de dietil sulfato (DES) / Induced mutagenesis in Solanum betaceum Cav. plants by the use of diethyl sulphate (DES)

RESUMEN La materia prima del fitomejoramiento es la variabilidad genética, que se presenta baja, en especies en proceso de domesticación, que no han sido sometidas a selección, como en Solanum betaceum. Una de las tecnologías para incrementar la variabilidad genética es la inducción de mutagénesis. El objetivo del estudio fue evaluar, a través de marcadores RAMs, las variaciones moleculares presentes en plántulas de S. betaceum, provenientes de semillas sometidas a diferentes concentraciones del agente mutante dietil sulfato (DES). Los loci polimórficos oscilaron entre 87,5 y 100 % y el número de alelos efectivos (Ne), entre 1,0 y 1,99. Los loci más polimórficos se observaron en TG, AG, ACA y CGA, que mostraron una heterosis media insesgada entre 0,34 y 0,51, que permite establecer que estos marcadores sean útiles para obtener mayor discriminación entre mutantes en S. betaceum. Las distancias genéticas oscilaron entre 0,30 y 1,0. El 81,28 % de estos registros se dieron entre 0,60 y 0,90; esto revela bajo nivel de cambios, debido al DES. Estos pequeños cambios contribuyeron a enriquecer la variabilidad genética de la muestra tratada con DES. Los marcadores RAMs fueron útiles para detectar cambios entre plantas provenientes de semillas tratadas con DES y plantas normales. La variabilidad genética entre tratamientos con DES fueron más altos que tratamientos sin DES. Las similitudes genéticas fueron bajas entre plantas tratadas y no tratadas y fueron altas, entre no tratadas. Los cambios producidos por DES fueron de baja magnitud; sin embargo, produjeron cambios en los niveles de variabilidad genética.

ABSTRACT The raw material for plant breeding is genetic variability, which is low in species in the process of domestication that have not been subjected to selection, as is the case with Solanum betaceum. One of the technologies to increase genetic variability is mutagenesis induction. The objective was to evaluate, through RAMs markers, the molecular variations present in S. betaceum seedlings from seeds previously subjected to different concentrations of the mutant agent diethyl sulfate (DES). The polymorphic loci ranged from 87.5 to 100%, number of effective alleles (Ne) between 1.0 and 1.99. The most polymorphic loci were observed in TG, AG, ACA, and CGA, which showed a mean unbiased heterosis between 0.34 and 0.51 with an average of 0.44, which allows establishing that these markers are useful to obtain greater discrimination between mutants in S. betaceum. Genetic distances ranged from 0.30 to 1.0. The 81.28% of these records were between 0.60 and 0.90. This reveals a low level of changes due to DES. These small changes contribute to enriching the genetic variability of the DES-treated sample. The RAMs markers were useful for detecting changes between plants from DES treated seeds and normal plants. Genetic variability between DES treatments was higher than non-DES treatments. Genetic similarities were low between treated and untreated plants and were high among untreated plants. The changes produced by DES were of low magnitude, however, they produced changes in the levels of genetic variability.

Evaluación del efecto protector contra la oxidación lipídica de fracciones obtenidas a partir del epicarpio de tomate de árbol (Solanum betaceum Sendtn) / Evaluation of protective effect against lipid oxidation of fractions obtained from tree tomato (Solanum betaceum Sendtn) epicarp / Avaliação do efeito protetor contra oxidação lipídica de frações obtidas a partir da casca de tomate da árvore (Solanum betaceum Sendtn)

Resumen Se obtuvieron fracciones a partir del epicarpio de tomate de árbol (Solanum betaceum Sendtn) y se evaluó su efecto protector contra la oxidación lipídica en carne de res cocida (CRC). Un extracto obtenido a través de extracción supercrítica fue fraccionado empleando, a su vez, extracción en fase sólida; dichas fracciones fueron adicionadas a CRC (concentración 200 mg/kg). El efecto protector fue determinado a través de la comparación de algunos productos de oxidación y el antioxidante sintético terbutilhidroxiquinona (TBHQ, concentración 200 mg/kg). Cuatro fracciones fueron obtenidas, la fracción de mayor polaridad mostró una alta eficiencia para inhibir la oxidación lipídica en CRC, reduciendo la formación de hidroperóxidosHPL y TBARS en 100 y 98,5%, respectivamente; esta mostró una eficiencia superior a la observada para el TBHQ. Esta fracción más activa fue sometida a análisis HPLC-DAD y se identificaron algunos compuestos fenólicos: tres catequinas (epigalocatequina, epicatequina y galato de epigalocatequina) y un ácido fenólico (rosmarínico). Los resultados obtenidos permitieron evidenciar que el epicarpio de tomate de árbol es fuente de antioxidantes con efecto protector sobre CRC, alternativa de aprovechamiento y valorización para dicho residuo agroindustrial.

Abstract Fractions from tree tomato (Solanum betaceum Sendtn) epicarp were obtained and their effect against lipid oxidation in cooked beef meat (CBM) was evaluated. An extract obtained by supercritical fluids extraction was fractionated using solid phase extraction and these fractions were added to CBM (at 300 mg/kg). The protector effect against lipid oxidation was determined by measuring some lipid oxidation products and comparison against the synthetic antioxidant tert-butylhydroquinone (TBHQ at 300 mg/ kg). Four fractions were obtained, the most polar fraction showed high efficiency to reduce the lipid oxidation on CBM, this fraction reduced the LHP and TBARS formation at 100.0 and 98.5%, respectively. This fraction showed an ability to retard lipid oxidation in CRC higher than TBHQ. The fraction with the highest protector effect was analyzed for its phenolic composition by High-Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD). Three catechin (epigallocatechin, epicatechin and epigallocatechin gallate) and a phenolic acid (rosmarinic acid) were identified. The results revealed that the tree tomato epicarp is a source of antioxidants with protective effect on CBM, an alternative for the use and valorization of this agroindustrial waste.

Resumo Frações da casca de tomate de árvore (Solanum betaceum Sendtn) foram obtidas e seu efeito para retardar a oxidação dos lipídios em carne bovina cozida (CBC) foi avaliado. Um extrato obtido através da extração supercrítica foi fracionado utilizando extração em fase sólida e as frações obtidas foram adicionadas á CBC (concentração 300 mg/kg). O efeito protetor contra a oxidação dos lipídios foi determinado medindo alguns produtos de oxidação e comparando com a terbutil-hidroquinona (TBHQ, concentração 300 mg/kg). Quatro frações foram obtidas, destas, a fração de maior polaridade mostrou a maior eficiência para inibir a oxidação dos lipídios da CBC, reduzindo a formação de HPL e TBARS em 100 e 98,5%, respectivamente; mostrou uma eficiência maior da observada para TBHQ. Esta foi submetida a análise por a cromatografia líquida de alta eficiência com detector de díodos (HPLC-DAD) e foram identificados alguns compostos fenólicos: três catequinas (epigalocatequina, galato de epicatequina e galato de epigalocatequina) e um ácido fenólico (rosmarínico). Os resultados permitiram evidênciar que a casca do tomate de árvore é uma fonte de compostos antioxidantes comefeito protetor sobre a CBC, esta é uma alternativa de utilização e aproveitamento deesse resíduo agroindustrial.

Efecto del consumo de jugo de tomate de árbol (Cyphomandra betacea) sobre el perfil lipídico y las concentraciones de glucosa en adultos con hiperlipidemia, ecuador / Effect of consumption of tree tomato juice (Cyphomandra betacea) on lipid profile and glucose concentrations in adults with hyperlipidemia, ecuador

En este trabajo se evaluó el efecto del consumo del jugo de tomate de árbol (Cyphomandra betacea) sobre parámetros nutricionales y bioquímicos en 54 voluntarios (44 mujeres y 10 hombres) con edades 45±8 años de Ecuador. Al inicio se les realizó una evaluación nutricional y bioquímica, luego se les invitó a consumir el jugo (100g de fruto en 150 ml de agua) diariamente durante 6 semanas. Finalizado este tiempo, se procedió a realizar la evaluación nutricional y bioquímica nuevamente. La evaluación nutricional determinó 67% de obesidad abdominal. Disminuyendo a 53% después del consumo del jugo de tomate de árbol. El 87% de los voluntarios antes del tratamiento, mostraron hipercolesterolemia; 40,7% y 46,3% tenían los triglicéridos y el LDL elevados y 18,5% valores de glucosa entre 98-130mg/dL. Valores que disminuyeron significativamente en este grupo después de la toma del jugo En general, tanto en voluntarios con y sin sobrepeso se observó disminución significativa de colesterol total, LDL y glucosa; sin cambios significativos en los valores de HDL. El consumo del jugo no afectó la actividad de las enzimas alanina-aminotransferasa ni aspartato-aminotransferasa, tampoco las concentraciones de creatinina, urea y ácido úrico; ni la presión arterial, demostrando que no afecta la función hepática ni renal. Estos resultados indican que el consumo del jugo de tomate de árbol, durante seis semanas, parece ejercer un efecto hipolipemiante y moduladora en el metabolismo de la glucosa y colocan a C betacea como uno de los frutos andinos con alto potencial nutraceutico. Sin embargo, estos aspectos deben ser investigados con más detalles(AU)

In this work the effect of consumption of tree tomato juice (Cyphomandra betacea) was evaluated on nutritional and biochemical parameters in 54 volunteers (44 women and 10 men) aged 45 ± 8 years-Ecuador. A nutritional and biochemical evaluation was performed in volunteers; then they were invited to drink tree tomato juice (100g of fruit in 150 ml of water) daily for 6 weeks. Finished these 6 weeks, volunteers were nutritional and biochemical evaluated again. 67% abdominal obesity was found, decreasing at 53% after drinking the established dosage of juice. 87% of the volunteers before treatment, showed hypercholesterolemia, 40.7% and 46.3% had hypertriglyceridemia and elevated LDL, respectively and 18.5% glucose concentrations between 98-130 mg/dL. These values decrease significantly in this group after drinking tomato juice. In general, Total Cholesterol, LDL and glucose concentrations decrease significantly after drinking tree tomato juice in all the voluntaries with or without overweight. There is no change in HDL concentrations. The consumption of tree tomato juice did not affect the activity of alanine aminotransferase or aspartate aminotransferase enzymes either creatinine, urea and uric acid concentrations, neither blood pressure suggesting that does not affect renal or liver function. These results indicate that consumption of tree tomato juice for six weeks appears to have a lipid-lowering and modulating effect on glucose metabolism, suggesting C betacea as one of the high Andean fruits nutraceutical potential. However, this issue should be investigated in more detail(AU)

Efecto de la ingesta de dietas con tomate (Lycopersicum esculentum) y tomate de árbol (Cyphomandra betacea (Cav) Sendtn) en los lípidos sanguíneos de ratas / Effect of dietary intake of tomato (Lycopersicum esculentum) and tree tomato (Cyphomandra betacea (Cav) Sendtn) on blood lipids in rats

Consumption of tomato (Lycopersicon esculentum ) and tree tomato ( Cyphomandra betacea ) is interesting for its nutritional composition and content of antioxidant compounds. This research evaluated the effect of diets intake with tomatoes on blood lipids in rats. First, fatty acids were determined in tomato seeds. Subsequently, rats (Sprague-Dawley, 6 females per group , 2 months old and weight 160 g) were fed with basal diet (AIN-93M) supplemented with 10% tomato and tomato tree at the expense of starch for 5 weeks. Efficiency and diet digestibility were calculated. Triglycerides (plasma), total cholesterol (serum), HDL (plasma) were measured and LDL was calculated. Results showed that seeds of C. betacea 7% had more polyunsaturated fatty acids than L. esculentum. Consumption of diets with both fruits decreased triglycerides and LDL and increased HDL. The results indicate the importance of encouraging the consumption of these fruits as a contribution to the prevention of some chronic diseases.

El consumo de tomate (Lycopersicum esculentum) y tomate de árbol (Cyphomandra betacea) resulta de interés por su composición nutricional y contenido de compuestos antioxidantes. Esta investigación evaluó el efecto de la ingesta de dietas con tomates en los lípidos sanguíneos de ratas. Primero, se determinaron los ácidos grasos en semillas de tomates. Posteriormente, a ratas (Sprague-Dawley, 6 hembras por grupo, 2 meses de edad y peso 160 g), se les suministró dieta basal (AIN- 93M), dietas suplementadas con 10% de tomate y de tomate de árbol a expensas del almidón, durante 5 semanas. Se calculó la eficiencia y digestibilidad de la dieta. Se midieron los triglicéridos (plasma), colesterol total (suero), HDL (plasma) y LDL calculado. Resultados indicaron 7% más de ácidos grasos poliinsaturados en semillas de C. betacea respecto a L. esculentum. El consumo de dietas con ambos frutos disminuyó los triglicéridos y LDL y aumentó el HDL. Los resultados señalan la importancia de incentivar el consumo de ambos frutos como una contribución a la prevención de algunas enfermedades crónicas.

Caracterización física, química y compuestos bioactivos de pulpa madura de tomate de árbol (Cyphomandra betacea) (Cav.) Sendtn.

El tomate de árbol (Cyphomandra betacea) es apreciado por sus cualidades nutritivas y ser fuente de compuestos antioxidantes, calcio, fósforo, potasio y hierro, azúcares, ácidos orgánicos, pectinas y flavonoides. En este estudio se determinaron parámetros físicos (peso, tamaño, fuerza compresión, humedad) y químicos (°Brix, acidez titulable, pH, proteína, fibra dietaria, cenizas, minerales y bioaccesibilidad de minerales, pectina, compuestos antioxidantes) del fruto procedente del Estado Aragua, Venezuela, como una contribución para incentivar y diversificar su consumo. La caracterización reflejó que los frutos estaban en estado de madurez para su consumo (°Brix 10,51, pH 3,5, acidez 0,02 g/100 ml y fuerza de compresión 4,32 Kgf/cm2), con rendimiento de pulpa del 74%. Los resultados del análisis de la pulpa madura reflejan un aporte de 30 Kcal/100g, fibra dietaria (4,10g/100g) y valores de fósforo, calcio, magnesio potasio y hierro de 331,32; 21,25; 21,18; 17,03; 7,44 mg/100g respectivamente. Se obtuvo 6,71 y 1,86 % de bioaccesibilidad para calcio y hierro. La pectina extraída (1,00 g/100g) resultó ser de alto metoxilo y alto grado de esterificación. La capacidad antioxidante de la pulpa madura (EC50 de 165 g /g DPPH y poder reductor de 0,07 mmol Fe +2/100g) pudiera atribuirse a la presencia de ácido ascórbico (23,32 mg/100g), licopeno (1,22 mg/100g), compuestos fenólicos (1,39 mg EAG/ g), antocianinas (0,29 mg cianidina/g) y taninos (0,40 mg catequina/ 100g). Los resultados obtenidos pueden ser utilizados para promover los beneficios sobre la salud y sugerir su uso como ingrediente funcional en el desarrollo de productos.

Physical, chemical and bioactive compounds of tree tomato (Cyphomandra betacea). Tree tomato (Cyphomandra betacea) is appreciated for its excellent nutritional qualities, being considered a good source of antioxidants compounds, calcium, phosphorus, potassium and iron, sugars, organic acids, pectins and flavonoids. In this study, were evaluated physical parameters (weight, size, compression strength and humidity) and chemical (°Brix, titratable acidity, pH, protein, dietary fiber, ash, minerals and their bioaccesibility, pectin, antioxidants compounds) of the fruit from the Aragua State, Venezuela, as a contribution to stimulate and diversify the consumption of the tree tomato. The characterization showed that the fruits were at the ripening stage for consumption (°Brix 10.51, pH 3.5, acidity 0.02 g/100ml and 4.32 Kgf/cm2 compression strength) gave a yield of 74% pulp. The analytical results of the ripped pulp showed a content of 30 Kcal/100g, dietary fiber (4.10g/100g), and minerals such as phosphorous, calcium, magnesium, potassium and iron (331.32, 21.25, 21.18, 17.03 and 7.44 mg/100g, respectively). Bioaccesibility values of 6.71 and 1.86% were reported for calcium and iron. The extracted pectin (1.00 g/100g) was classified as high methoxyl with high degree of esterification. The antioxidant capacity of the ripped pulp (EC50 of 165.00 g /g DPPH and reducing power of 0.07 mmol Fe +2/100g), could be attributed to the presence of ascorbic acid (23.32 mg/100g), lycopene (1.22 mg/100 g), and phenolic compounds (1.39 mg GAE/g), anthocyanins (0.29 mg cyanidin/ g) and tannins (0.40 mg catechin/100g).The results obtained encourage the nutritional benefits and suggest applications as a functional ingredient in food product development.

Genetic variation in the Solanaceae fruit bearing species lulo and tree tomato revealed by Conserved Ortholog (COSII) markers

The Lulo or naranjilla (Solanum quitoense Lam.) and the tree tomato or tamarillo (Solanum betaceum Cav. Sendt.) are both Andean tropical fruit species with high nutritional value and the potential for becoming premium products in local and export markets. Herein, we present a report on the genetic characterization of 62 accessions of lulos (n = 32) and tree tomatoes (n = 30) through the use of PCR-based markers developed from single-copy conserved orthologous genes (COSII) in other Solanaceae (Asterid) species. We successfully PCR amplified a set of these markers for lulos (34 out of 46 initially tested) and tree tomatoes (26 out of 41) for molecular studies. Six polymorphic COSII markers were found in lulo with a total of 47 alleles and five polymorphic markers in tree tomato with a total of 39 alleles in the two populations. Further genetic analyses indicated a high population structure (with F ST > 0.90), which may be a result of low migration between populations, adaptation to various niches and the number of markers evaluated. We propose COSII markers as sound tools for molecular studies, conservation and the breeding of these two fruit species.